LOCUS       sequence-248076-       16709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible B4GALT1 expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16709)
  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 16709)
  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: B4GALT1-Dox
FEATURES             Location/Qualifiers
     source          1..16709
                     /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(1569..1593)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1748..1803
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2164..2180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2188..2204
                     /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(2212..2218,2219..2236,2237..2242))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2257..2278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    2402..2427
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    3099..3119
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        3187..3398
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          3399..4337
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    4362..4386
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4396..5103
                     /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(5106..5130)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5285..5340
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5701..5717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5725..5741
                     /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(5749..5755,5756..5773,5774..5779))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5794..5815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    5939..5964
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    6636..6656
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        6681..7859
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6912..7850
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    7876..7900
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      7901..7910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7907..8620
                     /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(8624..8648)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    8803..8858
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9219..9235)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9243..9259
                     /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(9267..9273,9274..9291,9292..9297))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9312..9333
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    10167..11003
                     /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(11113..11132)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             11124..11933
                     /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(11191..11210)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(12186..12774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12266..12285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12945..13736,13737..13805))
                     /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     13568..13587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13806..13910)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(13936..14391)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14073..14094)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14285..14304
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14518..14540
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14532..14549
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14533..14549
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14559..14578
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14559..14577
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    14623..14643
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    14661..14698
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     protein_bind    14708..14732
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             14748..15347
                     /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(14748..14767)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     15244..15264
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(15349..15373)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    15528..15583
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     misc_feature    16013..16038
                     /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 ccaccatgag gtttctgagg ccggtcctgg gcggcagcgc
      421 cgcgatgccg ggcgcgaccc tgcagcgggc ctgccgcctg ctcgtggccg tctgcgcgct
      481 gcacctcggc gtcaccctgg tctattacct ctcgggccgc gacctgagcc gcctgcctca
      541 gctagtcggg gtttcctcta cacttcggag cggcacgatc ggcgccaccg ccaacaagca
      601 gccccccggc gcgcggccgc cgcctcctgt gggagtctcc tcgaagcctc gcccgggtcc
      661 cgactccagc cctggtacgg cttttgaccc cggcctgaag agcaactgga cttcggtcct
      721 agtgccccct accactgcac tgttgacgct gcccgcttgc cccgaggagt ccccgctgct
      781 cgttggcccc atggtgattg actttaatat tgctgtggac ctggagcttt tggcaaagaa
      841 gaacccagag ataaagatgg gcggtcgtta ttcccccaag gactgtattt ctcctcacaa
      901 ggtggccatc attatcccat ttcgcaaccg gcaggagcac ctcaaatatt ggctgtatta
      961 tttgcatccg gtcctacagc gccagcaact ggactatggt atctacgtca tcaaccaggc
     1021 tggagacacc atgttcaatc gagctaagct tctcaacata ggcttccaag aggccttgaa
     1081 ggaccatgac tacaactgct ttgtgttcag tgatgtggac cttattccaa tggacgacca
     1141 taatgcctac aggtgttttt cacagccccg gcacatttct gttgcaatgg acaagtttgg
     1201 gtttagcctg ccttatgttc agtactttgg aggcgtttct gccctcagta aacaacagtt
     1261 tcttgccatc aatggatttc ctaataatta ttggggctgg ggaggagaag atgatgacat
     1321 ttttaacaga atagttcata agggaatgtc tatatcacgc ccaaatgctg tagttggaag
     1381 atgtcgcatg atacgacact caagagacaa gaaaaatgag cccaatcctc agaggtttga
     1441 ccggattgct cacacaaagg aaactatgcg ctttgatggt ttgaactcac ttacctacca
     1501 ggtgttgaac gtagagagat acccattata taccaaaatc acagtggaca tcgggacacc
     1561 aagatagtac ccagctttct tgtacaaagt ggtacgcgtg aattcactcc tcaggtgcag
     1621 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     1681 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
     1741 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     1801 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
     1861 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
     1921 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     1981 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     2041 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     2101 atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc
     2161 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2221 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2281 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatcgacag
     2341 tactaagctt ggtgcgtttt tatgcttgta gtattgtata atgtttttaa gatccaagct
     2401 tctgacctct tctcttcctc ccacagggcc tcgagacaat tgcatcgatg gtaccagtga
     2461 aagtctgcaa tgaattcgag ttggcgcgcc tgtcattcta aatctctctt tcagcctaaa
     2521 gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag
     2581 aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc
     2641 ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct
     2701 agcgggggag ggacgtaatt acatccctgg gggctttggg agggggctgt ccccgtgagc
     2761 tcccaggcgc gcctgtcatt ctaaatctct ctttcagcct aaagcttttt ccccgtatcc
     2821 ccccaggtgt ctgcaggctc aaagagcagc gagaagcgtt cagaggaaag cgatcccgtg
     2881 ccaccttccc cgtgcccggg ctgtccccgc acgctgccgg ctcggggatg cggggggagc
     2941 gccggaccgg agcggagccc cgggcggctc gctgctgccc cctagcgggg gagggacgta
     3001 attacatccc tgggggcttt gggggggggc tgtccccgtg agctcccagg cgcgccaact
     3061 cggtaccgta tcgatgtcga cgttaacgct agtgatatca actttgtata gaaaagttgg
     3121 ctccgaattc gcccttgaca gcagagatcc agtttggtta attaattggc tccggtgccc
     3181 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgtgggga ggggtcggca
     3241 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3301 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     3361 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
     3421 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
     3481 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
     3541 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     3601 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     3661 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     3721 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     3781 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     3841 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     3901 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     3961 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     4021 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     4081 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     4141 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     4201 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     4261 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     4321 tttttcttcc atttcaggtg gatcccaagg gcgaattcga cccaagtttg tacaaaaaag
     4381 caggctgatg ccaccatgtc taggctggac aagagcaaag tcataaacgg agctctggaa
     4441 ttactcaatg gtgtcggtat cgaaggcctg acgacaagga aactcgctca aaagctggga
     4501 gttgagcagc ctaccctgta ctggcacgtg aagaacaagc gggccctgct cgatgccctg
     4561 ccaatcgaga tgctggacag gcatcatacc cacttctgcc ccctggaagg cgagtcatgg
     4621 caagactttc tgcggaacaa cgccaagtca taccgctgtg ctctcctctc acatcgcgac
     4681 ggggctaaag tgcatctcgg cacccgccca acagagaaac agtacgaaac cctggaaaat
     4741 cagctcgcgt tcctgtgtca gcaaggcttc tccctggaga acgcactgta cgctctgtcc
     4801 gccgtgggcc actttacact gggctgcgta ttggaggaac aggagcatca agtagcaaaa
     4861 gaggaaagag agacacctac caccgattct atgcccccac ttctgagaca agcaattgag
     4921 ctgttcgacc ggcagggagc cgaacctgcc ttccttttcg gcctggaact aatcatatgt
     4981 ggcctggaga aacagctaaa gtgcgaaagc ggcgggccga ccgacgccct tgacgatttt
     5041 gacttagaca tgctcccagc cgatgccctt gacgattttg accttgacat gctccccggg
     5101 taaataccca gctttcttgt acaaagtggt acgcgtgaat tcactcctca ggtgcaggct
     5161 gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat
     5221 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     5281 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     5341 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     5401 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
     5461 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     5521 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     5581 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     5641 gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg
     5701 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5761 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5821 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tcgacagtac
     5881 taagcttggt ctaataccca atctctcgtc ttatccagat gttttatacg ccaagcttct
     5941 gacctcttct cttcctccca cagggcctcg agacaattgc atcgatggta ccagtgaaag
     6001 tctgcaatga attcgagttg gcgcgcctgt cattctaaat ctctctttca gcctaaagct
     6061 ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg
     6121 aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg
     6181 gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc
     6241 gggggaggga cgtaattaca tccctggggg ctttgggagg gggctgtccc cgtgagctcc
     6301 caggcgcgcc tgtcattcta aatctctctt tcagcctaaa gctttttccc cgtatccccc
     6361 caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca
     6421 ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc
     6481 ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt
     6541 acatccctgg gggctttggg ggggggctgt ccccgtgagc tcccaggcgc gccaactcgg
     6601 taccgtatcg atgtcgacgt taacgctagt gatatcaact ttgtatagaa aagttggctc
     6661 cgaattcgcc cttattaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca
     6721 cagtccccga gaagttgtgg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc
     6781 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     6841 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     6901 ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg
     6961 gcccttgcgt gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc
     7021 ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc
     7081 gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc
     7141 ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg
     7201 ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg
     7261 gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt
     7321 cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg
     7381 gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa
     7441 ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg
     7501 cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac
     7561 aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg
     7621 cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg
     7681 gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc
     7741 cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt
     7801 tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag
     7861 aagggcgaat tcgacccaag tttgtacaaa aaagcaggct gccaccatgg tgtctaaggg
     7921 cgaggagctg attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa
     7981 ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat
     8041 gagaatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag
     8101 cttcatgtac ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa
     8161 gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt
     8221 gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat
     8281 cagaggggtg aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga
     8341 ggcctccacc gagatgctgt accccgctga cggcggcctg gaaggcagaa gcgacatggc
     8401 cctgaagctc gtgggcgggg gccacctgat ctgcaacttg aagaccacat acagatccaa
     8461 gaaacccgct aagaacctca agatgcccgg cgtctactat gtggacagaa gactggaaag
     8521 aatcaaggag gccgacaaag agacatacgt cgagcagcac gaggtggctg tggccagata
     8581 ctgcgacctc cctagcaaac tggggcacaa acttaattga taaacccagc tttcttgtac
     8641 aaagtggtac gcgtgaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg
     8701 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     8761 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     8821 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     8881 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     8941 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     9001 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     9061 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     9121 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     9181 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     9241 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     9301 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     9361 cagtcgggaa acctgtcgtg ccagcggatc gacagtacta agcttggtga attcccttat
     9421 gtgagtgtaa aaggcaggcg agtttgtccc caattgcatc gatggtacca gtgaaagtct
     9481 gcaatgaatt cgagttggcg cgcctgtcat tctaaatctc tctttcagcc taaagctttt
     9541 tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa
     9601 gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat
     9661 gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg
     9721 ggagggacgt aattacatcc ctgggggctt tgggaggggg ctgtccccgt gagctcccag
     9781 gcgcgcctgt cattctaaat ctctctttca gcctaaagct ttttccccgt atccccccag
     9841 gtgtctgcag gctcaaagag cagcgagaag cgttcagagg aaagcgatcc cgtgccacct
     9901 tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg gatgcggggg gagcgccgga
     9961 ccggagcgga gccccgggcg gctcgctgct gccccctagc gggggaggga cgtaattaca
    10021 tccctggggg ctttgggggg gggctgtccc cgtgagctcc caggcgcgcc aactcggtac
    10081 cgtatcgatg tcgacggtgc gtttttatgc ttgtagtatt gtataatgtt tttaagatcc
    10141 tagggataac agggtaatcg gccgctacta gggacaggat tggtgacaga aaagccccat
    10201 ccttaggcct cctccttcct agtctcctga tattgggtct aacccccacc tcctgttagg
    10261 cagattcctt atctggtgac acacccccat ttcctggagc catctctctc cttgccagaa
    10321 cctctaaggt ttgcttacga tggagccaga gaggatcctg ggagggagag cttggcaggg
    10381 ggtgggaggg aaggggggga tgcgtgacct gcccggttct cagtggccac cctgcgctac
    10441 cctctcccag aacctgagct gctctgacgc ggctgtctgg tgcgtttcac tgatcctggt
    10501 gctgcagctt ccttacactt cccaagagga gaagcagttt ggaaaaacaa aatcagaata
    10561 agttggtcct gagttctaac tttggctctt cacctttcta gtccccaatt tatattgttc
    10621 ctccgtgcgt cagttttacc tgtgagataa ggccagtagc cagccccgtc ctggcagggc
    10681 tgtggtgagg aggggggtgt ccgtgtggaa aactcccttt gtgagaatgg tgcgtcctag
    10741 gtgttcacca ggtcgtggcc gcctctactc cctttctctt tctccatcct tctttcctta
    10801 aagagtcccc agtgctatct gggacatatt cctccgccca gagcagggtc ccgcttccct
    10861 aaggccctgc tctgggcttc tgggtttgag tccttggcaa gcccaggaga ggcgctcagg
    10921 cttccctgtc ccccttcctc gtccaccatc tcatgcccct ggctctcctg ccccttccct
    10981 acaggggttc ctggctctgc tctccaccgc gggagctctg gcccgtgtct caaaatctct
    11041 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
    11101 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
    11161 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
    11221 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
    11281 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
    11341 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
    11401 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
    11461 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
    11521 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
    11581 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
    11641 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
    11701 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
    11761 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
    11821 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
    11881 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
    11941 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
    12001 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gacggtatca
    12061 ctagtattac agaggtaagt tataacagtc gcctaaccca attgagggga taacgcagga
    12121 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
    12181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
    12241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
    12301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
    12361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
    12421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
    12481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
    12541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
    12601 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
    12661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
    12721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
    12781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
    12841 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
    12901 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
    12961 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
    13021 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
    13081 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
    13141 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
    13201 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
    13261 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
    13321 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
    13381 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
    13441 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
    13501 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    13561 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    13621 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    13681 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    13741 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    13801 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
    13861 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
    13921 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
    13981 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
    14041 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
    14101 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
    14161 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
    14221 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
    14281 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
    14341 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
    14401 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
    14461 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
    14521 agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg
    14581 aattgggtac cgtatcgatg tcgacgttaa cgctagtgat atcaactttg tatagaaaag
    14641 ttggctccga atttctcgag ggcttgtcga cgacggcgga ctccgtcgtc aggatcatga
    14701 attcgaccca agtttgtaca aaaaagcagg ctgaacggcc tgccaccatg accgagtaca
    14761 agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg
    14821 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc
    14881 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
    14941 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
    15001 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
    15061 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
    15121 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
    15181 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
    15241 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
    15301 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatac ccagctttct
    15361 tgtacaaagt ggtacgcgtg aattcactcc tcaggtgcag gctgcctatc agaaggtggt
    15421 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
    15481 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
    15541 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
    15601 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
    15661 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
    15721 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
    15781 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
    15841 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
    15901 atggagatcc ctcgacctgc agcccaagct tgaattctct agaattaccc tgttatccct
    15961 aggtttaccg agctcttatt ggttttcaaa cttcattgac tgtgccaagc ttctgacctc
    16021 ttctcttcct cccacagggc ctcgagacaa ttgcatcgat ggtaccagtg aaagtctgca
    16081 atgaattcga gttggcgcgc ctgtcattct aaatctctct ttcagcctaa agctttttcc
    16141 ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg
    16201 atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg
    16261 gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga
    16321 gggacgtaat tacatccctg ggggctttgg gagggggctg tccccgtgag ctcccaggcg
    16381 cgcctgtcat tctaaatctc tctttcagcc taaagctttt tccccgtatc cccccaggtg
    16441 tctgcaggct caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt gccaccttcc
    16501 ccgtgcccgg gctgtccccg cacgctgccg gctcggggat gcggggggag cgccggaccg
    16561 gagcggagcc ccgggcggct cgctgctgcc ccctagcggg ggagggacgt aattacatcc
    16621 ctgggggctt tggggggggg ctgtccccgt gagctcccag gcgcgccaac tcggtaccgt
    16681 atcgatgtcg acgttaacgc tagtgatat
//
