LOCUS       sequence-248079-       17231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible FUT8 expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17231)
  AUTHORS   Chang MM, Gaidukov L, Jung G, Tseng WA, Scarcelli JJ, Cornell R, 
            Marshall JK, Lyles JL, Sakorafas P, Chu AA, Cote K, Tzvetkova B, 
            Dolatshahi S, Sumit M, Mulukutla BC, Lauffenburger DA, Figueroa B 
            Jr, Summers NM, Lu TK, Weiss R
  TITLE     Small-molecule control of antibody N-glycosylation in engineered 
            mammalian cells.
  JOURNAL   Nat Chem Biol. 2019 May 20. pii: 10.1038/s41589-019-0288-4. doi: 
            10.1038/s41589-019-0288-4.
  PUBMED    31110306
REFERENCE   2  (bases 1 to 17231)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2019 May 20. pii: 10.1038/s41589-019-0288-4. doi: 
            10.1038/s41589-019-0288-4."
COMMENT     Sequence Label: FUT8-Dox
FEATURES             Location/Qualifiers
     source          1..17231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..21
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    66..84
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    101..119
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    137..155
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    173..191
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    208..226
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    244..262
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        274..312
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     309..333
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    352..376
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2115..2139)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2294..2349
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2710..2726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2734..2750
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2758..2764,2765..2782,2783..2788))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2803..2824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    2948..2973
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    3645..3665
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        3709..3920
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          3921..4859
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    4884..4908
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4918..5625
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     protein_bind    complement(5628..5652)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5807..5862
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6223..6239)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6247..6263
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6271..6277,6278..6295,6296..6301))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6316..6337
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    6461..6486
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    7158..7178
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        7203..8381
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7434..8372
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    8398..8422
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      8423..8432
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8429..9142
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     protein_bind    complement(9146..9170)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    9325..9380
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9741..9757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9765..9781
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(9789..9795,9796..9813,9814..9819))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9834..9855
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    10689..11525
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(11635..11654)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             11646..12455
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(11713..11732)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(12708..13296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12788..12807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13467..14258,14259..14327))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     14090..14109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14328..14432)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(14458..14913)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14595..14616)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14807..14826
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15040..15062
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15054..15071
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15055..15071
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15081..15100
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        15081..15099
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    15145..15165
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    15183..15220
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     protein_bind    15230..15254
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             15270..15869
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(15270..15289)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     15766..15786
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(15871..15895)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    16050..16105
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     misc_feature    16535..16560
                     /label=SA
                     /note="splice acceptor site"
ORIGIN
        1 caactttgta tagaaaagtt ggctccgaat tcgcccttca ggtccgaggt tctagacgag
       61 tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag tgatagagaa
      121 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc
      181 agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt atgtcgagtt
      241 tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg ggaggcctat
      301 ataagcagag ctcgtttagt gaaccgtcag atcgcaaagg gcgaattcga cccaagtttg
      361 tacaaaaaag caggctgatg ccaccatgcg ggcatggact ggttcctggc gttggattat
      421 gctcattctt tttgcctggg ggaccttatt gttttatata ggtggtcatt tggttcgaga
      481 taatgaccac cctgaccatt ctagcagaga actctccaag attcttgcaa agctggagcg
      541 cttaaaacaa caaaatgaag acttgaggag aatggctgag tctctccgaa taccagaagg
      601 ccctattgat caggggacag ctacaggaag agtccgtgtt ttagaagaac agcttgttaa
      661 ggccaaagaa cagattgaaa attacaagaa acaagctagg aatgatctgg gaaaggatca
      721 tgaaatctta aggaggagga ttgaaaatgg agctaaagag ctctggtttt ttctacaaag
      781 tgaattgaag aaattaaaga aattagaagg aaacgaactc caaagacatg cagatgaaat
      841 tcttttggat ttaggacatc atgaaaggtc tatcatgaca gatctatact acctcagtca
      901 aacagatgga gcaggtgagt ggcgggaaaa agaagccaaa gatctgacag agctggtcca
      961 gcggagaata acatatctgc agaatcccaa ggactgcagc aaagccagaa agctggtatg
     1021 taatatcaac aaaggctgtg gctatggatg tcaactccat catgtggttt actgcttcat
     1081 gattgcttat ggcacccagc gaacactcat cttggaatct cagaattggc gctatgctac
     1141 tggaggatgg gagactgtgt ttagacctgt aagtgagaca tgcacagaca ggtctggcct
     1201 ctccactgga cactggtcag gtgaagtgaa ggacaaaaat gttcaagtgg tcgagctccc
     1261 cattgtagac agcctccatc ctcgtcctcc ttacttaccc ttggctgtac cagaagacct
     1321 tgcagatcga ctcctgagag tccatggtga tcctgcagtg tggtgggtat cccagtttgt
     1381 caaatacttg atccgtccac aaccttggct ggaaagggaa atagaagaaa ccaccaagaa
     1441 gcttggcttc aaacatccag ttattggagt ccatgtcaga cgcactgaca aagtgggaac
     1501 agaagcagcc ttccatccca ttgaggaata catggtacac gttgaagaac attttcagct
     1561 tctcgaacgc agaatgaaag tggataaaaa aagagtgtat ctggccactg atgacccttc
     1621 tttgttaaag gaggcaaaga caaagtactc caattatgaa tttattagtg ataactctat
     1681 ttcttggtca gctggactac acaaccgata cacagaaaat tcacttcggg gcgtgatcct
     1741 ggatatacac tttctctccc aggctgactt ccttgtgtgt actttttcat cccaggtctg
     1801 tagggttgct tatgaaatca tgcaaacact gcatcctgat gcctctgcaa acttccattc
     1861 tttagatgac atctactatt ttggaggcca aaatgcccac aaccagattg cagtttatcc
     1921 tcaccaacct cgaactaaag aggaaatccc catggaacct ggagatatca ttggtgtggc
     1981 tggaaaccat tggaatggtt actctaaagg tgtcaacaga aaactaggaa aaacaggcct
     2041 gtacccttcc tacaaagtcc gagagaagat agaaacagtc aaatacccta catatcctga
     2101 agctgaaaaa tagtacccag ctttcttgta caaagtggta cgcgtgaatt cactcctcag
     2161 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     2221 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     2281 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     2341 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     2401 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     2461 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     2521 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     2581 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     2641 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc
     2701 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2761 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2821 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     2881 cgacagtact aagcttggtg cgtttttatg cttgtagtat tgtataatgt ttttaagatc
     2941 caagcttctg acctcttctc ttcctcccac agggcctcga gacaattgca tcgatggtac
     3001 cagtgaaagt ctgcaatgaa ttcgagttgg cgcgcctgtc attctaaatc tctctttcag
     3061 cctaaagctt tttccccgta tccccccagg tgtctgcagg ctcaaagagc agcgagaagc
     3121 gttcagagga aagcgatccc gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc
     3181 cggctcgggg atgcgggggg agcgccggac cggagcggag ccccgggcgg ctcgctgctg
     3241 ccccctagcg ggggagggac gtaattacat ccctgggggc tttgggaggg ggctgtcccc
     3301 gtgagctccc aggcgcgcct gtcattctaa atctctcttt cagcctaaag ctttttcccc
     3361 gtatcccccc aggtgtctgc aggctcaaag agcagcgaga agcgttcaga ggaaagcgat
     3421 cccgtgccac cttccccgtg cccgggctgt ccccgcacgc tgccggctcg gggatgcggg
     3481 gggagcgccg gaccggagcg gagccccggg cggctcgctg ctgcccccta gcgggggagg
     3541 gacgtaatta catccctggg ggctttgggg gggggctgtc cccgtgagct cccaggcgcg
     3601 ccaactcggt accgtatcga tgtcgacgtt aacgctagtg atatcaactt tgtatagaaa
     3661 agttggctcc gaattcgccc ttattaattg gctccggtgc ccgtcagtgg gcagagcgca
     3721 catcgcccac agtccccgag aagttgtggg gaggggtcgg caattgaacc ggtgcctaga
     3781 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     3841 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     3901 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
     3961 cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg
     4021 atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc
     4081 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct
     4141 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt
     4201 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc
     4261 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc
     4321 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt
     4381 ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct
     4441 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg
     4501 gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt
     4561 cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg
     4621 agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt
     4681 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg
     4741 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg
     4801 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg
     4861 tggatcccaa gggcgaattc gacccaagtt tgtacaaaaa agcaggctga tgccaccatg
     4921 tctaggctgg acaagagcaa agtcataaac ggagctctgg aattactcaa tggtgtcggt
     4981 atcgaaggcc tgacgacaag gaaactcgct caaaagctgg gagttgagca gcctaccctg
     5041 tactggcacg tgaagaacaa gcgggccctg ctcgatgccc tgccaatcga gatgctggac
     5101 aggcatcata cccacttctg ccccctggaa ggcgagtcat ggcaagactt tctgcggaac
     5161 aacgccaagt cataccgctg tgctctcctc tcacatcgcg acggggctaa agtgcatctc
     5221 ggcacccgcc caacagagaa acagtacgaa accctggaaa atcagctcgc gttcctgtgt
     5281 cagcaaggct tctccctgga gaacgcactg tacgctctgt ccgccgtggg ccactttaca
     5341 ctgggctgcg tattggagga acaggagcat caagtagcaa aagaggaaag agagacacct
     5401 accaccgatt ctatgccccc acttctgaga caagcaattg agctgttcga ccggcaggga
     5461 gccgaacctg ccttcctttt cggcctggaa ctaatcatat gtggcctgga gaaacagcta
     5521 aagtgcgaaa gcggcgggcc gaccgacgcc cttgacgatt ttgacttaga catgctccca
     5581 gccgatgccc ttgacgattt tgaccttgac atgctccccg ggtaaatacc cagctttctt
     5641 gtacaaagtg gtacgcgtga attcactcct caggtgcagg ctgcctatca gaaggtggtg
     5701 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
     5761 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     5821 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     5881 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     5941 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     6001 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     6061 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     6121 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     6181 tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
     6241 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6301 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6361 tttccagtcg ggaaacctgt cgtgccagcg gatcgacagt actaagcttg gtctaatacc
     6421 caatctctcg tcttatccag atgttttata cgccaagctt ctgacctctt ctcttcctcc
     6481 cacagggcct cgagacaatt gcatcgatgg taccagtgaa agtctgcaat gaattcgagt
     6541 tggcgcgcct gtcattctaa atctctcttt cagcctaaag ctttttcccc gtatcccccc
     6601 aggtgtctgc aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac
     6661 cttccccgtg cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg
     6721 gaccggagcg gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta
     6781 catccctggg ggctttggga gggggctgtc cccgtgagct cccaggcgcg cctgtcattc
     6841 taaatctctc tttcagccta aagctttttc cccgtatccc cccaggtgtc tgcaggctca
     6901 aagagcagcg agaagcgttc agaggaaagc gatcccgtgc caccttcccc gtgcccgggc
     6961 tgtccccgca cgctgccggc tcggggatgc ggggggagcg ccggaccgga gcggagcccc
     7021 gggcggctcg ctgctgcccc ctagcggggg agggacgtaa ttacatccct gggggctttg
     7081 ggggggggct gtccccgtga gctcccaggc gcgccaactc ggtaccgtat cgatgtcgac
     7141 gttaacgcta gtgatatcaa ctttgtatag aaaagttggc tccgaattcg cccttattaa
     7201 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgt
     7261 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     7321 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     7381 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     7441 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     7501 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     7561 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     7621 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     7681 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     7741 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     7801 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     7861 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     7921 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     7981 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     8041 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     8101 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     8161 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     8221 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     8281 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     8341 cagtggttca aagttttttt cttccatttc aggtgtcgtg agaagggcga attcgaccca
     8401 agtttgtaca aaaaagcagg ctgccaccat ggtgtctaag ggcgaggagc tgattaagga
     8461 gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac
     8521 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga
     8581 gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcaa
     8641 aaccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct tccctgaggg
     8701 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     8761 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttccc
     8821 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggcctcca ccgagatgct
     8881 gtaccccgct gacggcggcc tggaaggcag aagcgacatg gccctgaagc tcgtgggcgg
     8941 gggccacctg atctgcaact tgaagaccac atacagatcc aagaaacccg ctaagaacct
     9001 caagatgccc ggcgtctact atgtggacag aagactggaa agaatcaagg aggccgacaa
     9061 agagacatac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc tccctagcaa
     9121 actggggcac aaacttaatt gataaaccca gctttcttgt acaaagtggt acgcgtgaat
     9181 tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     9241 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     9301 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     9361 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     9421 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     9481 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     9541 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     9601 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     9661 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc
     9721 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     9781 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     9841 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     9901 tgccagcgga tcgacagtac taagcttggt gaattccctt atgtgagtgt aaaaggcagg
     9961 cgagtttgtc cccaattgca tcgatggtac cagtgaaagt ctgcaatgaa ttcgagttgg
    10021 cgcgcctgtc attctaaatc tctctttcag cctaaagctt tttccccgta tccccccagg
    10081 tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt
    10141 ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg agcgccggac
    10201 cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac gtaattacat
    10261 ccctgggggc tttgggaggg ggctgtcccc gtgagctccc aggcgcgcct gtcattctaa
    10321 atctctcttt cagcctaaag ctttttcccc gtatcccccc aggtgtctgc aggctcaaag
    10381 agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt
    10441 ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg
    10501 cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg
    10561 gggggctgtc cccgtgagct cccaggcgcg ccaactcggt accgtatcga tgtcgacggt
    10621 gcgtttttat gcttgtagta ttgtataatg tttttaagat cctagggata acagggtaat
    10681 cggccgctac tagggacagg attggtgaca gaaaagcccc atccttaggc ctcctccttc
    10741 ctagtctcct gatattgggt ctaaccccca cctcctgtta ggcagattcc ttatctggtg
    10801 acacaccccc atttcctgga gccatctctc tccttgccag aacctctaag gtttgcttac
    10861 gatggagcca gagaggatcc tgggagggag agcttggcag ggggtgggag ggaagggggg
    10921 gatgcgtgac ctgcccggtt ctcagtggcc accctgcgct accctctccc agaacctgag
    10981 ctgctctgac gcggctgtct ggtgcgtttc actgatcctg gtgctgcagc ttccttacac
    11041 ttcccaagag gagaagcagt ttggaaaaac aaaatcagaa taagttggtc ctgagttcta
    11101 actttggctc ttcacctttc tagtccccaa tttatattgt tcctccgtgc gtcagtttta
    11161 cctgtgagat aaggccagta gccagccccg tcctggcagg gctgtggtga ggaggggggt
    11221 gtccgtgtgg aaaactccct ttgtgagaat ggtgcgtcct aggtgttcac caggtcgtgg
    11281 ccgcctctac tccctttctc tttctccatc cttctttcct taaagagtcc ccagtgctat
    11341 ctgggacata ttcctccgcc cagagcaggg tcccgcttcc ctaaggccct gctctgggct
    11401 tctgggtttg agtccttggc aagcccagga gaggcgctca ggcttccctg tcccccttcc
    11461 tcgtccacca tctcatgccc ctggctctcc tgccccttcc ctacaggggt tcctggctct
    11521 gctctccacc gcgggagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
    11581 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
    11641 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
    11701 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
    11761 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
    11821 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
    11881 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
    11941 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
    12001 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
    12061 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
    12121 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
    12181 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
    12241 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
    12301 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
    12361 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
    12421 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
    12481 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
    12541 acgtgagtta cgcgtcgttc cactgagcgt cagacggtat cactagtatt acagaggtaa
    12601 gttataacag tcgcctaacc caattgaggg gataacgcag gaaagaacat gtgagcaaaa
    12661 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
    12721 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
    12781 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
    12841 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
    12901 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
    12961 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
    13021 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    13081 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    13141 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    13201 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
    13261 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
    13321 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
    13381 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
    13441 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
    13501 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
    13561 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
    13621 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
    13681 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
    13741 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
    13801 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
    13861 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
    13921 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
    13981 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
    14041 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
    14101 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
    14161 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
    14221 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
    14281 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
    14341 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
    14401 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
    14461 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
    14521 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
    14581 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
    14641 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
    14701 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
    14761 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
    14821 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
    14881 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
    14941 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
    15001 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
    15061 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgtatcga
    15121 tgtcgacgtt aacgctagtg atatcaactt tgtatagaaa agttggctcc gaatttctcg
    15181 agggcttgtc gacgacggcg gactccgtcg tcaggatcat gaattcgacc caagtttgta
    15241 caaaaaagca ggctgaacgg cctgccacca tgaccgagta caagcccacg gtgcgcctcg
    15301 ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc
    15361 ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag
    15421 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
    15481 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
    15541 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
    15601 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
    15661 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
    15721 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
    15781 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
    15841 ggtgcatgac ccgcaagccc ggtgcctgat acccagcttt cttgtacaaa gtggtacgcg
    15901 tgaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
    15961 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
    16021 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
    16081 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
    16141 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
    16201 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
    16261 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
    16321 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
    16381 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct
    16441 gcagcccaag cttgaattct ctagaattac cctgttatcc ctaggtttac cgagctctta
    16501 ttggttttca aacttcattg actgtgccaa gcttctgacc tcttctcttc ctcccacagg
    16561 gcctcgagac aattgcatcg atggtaccag tgaaagtctg caatgaattc gagttggcgc
    16621 gcctgtcatt ctaaatctct ctttcagcct aaagcttttt ccccgtatcc ccccaggtgt
    16681 ctgcaggctc aaagagcagc gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc
    16741 cgtgcccggg ctgtccccgc acgctgccgg ctcggggatg cggggggagc gccggaccgg
    16801 agcggagccc cgggcggctc gctgctgccc cctagcgggg gagggacgta attacatccc
    16861 tgggggcttt gggagggggc tgtccccgtg agctcccagg cgcgcctgtc attctaaatc
    16921 tctctttcag cctaaagctt tttccccgta tccccccagg tgtctgcagg ctcaaagagc
    16981 agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccc gggctgtccc
    17041 cgcacgctgc cggctcgggg atgcgggggg agcgccggac cggagcggag ccccgggcgg
    17101 ctcgctgctg ccccctagcg ggggagggac gtaattacat ccctgggggc tttggggggg
    17161 ggctgtcccc gtgagctccc aggcgcgcca actcggtacc gtatcgatgt cgacgttaac
    17221 gctagtgata t
//
