LOCUS       sequence-232732-       12235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For targeted integration and inducible expression of a human NODAL 
            splice variant (with an N-terminal MYC tag on the mature peptide) 
            using doxycycline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12235)
  AUTHORS   Findlay SD, Bilyk O, Lypka K, Waskiewicz AJ, Postovit L
  TITLE     Genetically regulated human NODAL splice variants are differentially
            post-transcriptionally processed and functionally distinct
  JOURNAL   (2018) bioRxiv 276170
REFERENCE   2  (bases 1 to 12235)
  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: "(2018) 
            bioRxiv 276170"
COMMENT     Sequence Label: AAVS1-TRE3G-NODALvar-matMYC
FEATURES             Location/Qualifiers
     source          1..12235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        34..412
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    42..60
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    78..96
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    114..132
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    150..168
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    186..204
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    222..240
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    258..276
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     321..345
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1138..1167
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     intron          1634..1699
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1829..1849
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2274..2395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2311..2330)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2365..2384
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2433..2657
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2646..2663
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2695..3294)
                     /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(2778..2798)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3275..3294
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(3304..3357)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    3381..3406
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    3413..4216
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(4255..4273)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4294..4310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4318..4334
                     /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(4342..4348,4349..4366,4367..4372))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4387..4408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4525..4542)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4696..5284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4776..4795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5408..5476,5477..6268)
                     /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     complement(5626..5645)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(6629..6648)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7243..7262
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(7665..7967)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(7767..7786)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     8090..8112
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8104..8121
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8105..8121
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8128..8147
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8128..8146
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    8181..9017
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    9091..9112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9127..9132,9133..9150,9151..9157)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9165..9181
                     /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)."
     primer_bind     9189..9205
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9548..9567
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    9566..9621
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     9667..9686
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(9777..10523)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(10561..10580)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          10588..11604
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(10642..10664)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        11882..12185
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttaataacta gttttcgtct tcaagaattc ctcgagttta ctccctatca gtgatagaga
       61 acgtatgaag agtttactcc ctatcagtga tagagaacgt atgcagactt tactccctat
      121 cagtgataga gaacgtataa ggagtttact ccctatcagt gatagagaac gtatgaccag
      181 tttactccct atcagtgata gagaacgtat ctacagttta ctccctatca gtgatagaga
      241 acgtatatcc agtttactcc ctatcagtga tagagaacgt ataagcttta ggcgtgtacg
      301 gtgggcgcct ataaaagcag agctcgttta gtgaaccgtc agatcgcctg gagcaattcc
      361 acaacacttt tgtcttatac caactttccg taccacttcc taccctcgta aagtcgaccc
      421 accatgcacg cccactgcct gcccttcctt ctgcacgcct ggtgggccct actccaggcg
      481 ggtgctgcga cggtggccac tgcgctcctg cgtacgcggg ggcagccctc gtcgccatcc
      541 cctctggcgt acatgctgag cctctaccgc gacccgctgc cgagggcaga catcatccgc
      601 agcctacagg cagaagatgt ggcagtggat gggcagaact ggacgtttgc ttttgacttc
      661 tccttcctga gccaacaaga ggatctggca tgggctgagc tccggctgca gctgtccagc
      721 cctgtggacc tccccactga gggctcactt gccattgaga ttttccacca gccaaagccc
      781 gacacagagc aggcttcaga cagctgctta gagcggtttc agatggacct attcactgtc
      841 actttgtccc aggtcacctt ttccttgggc agcatggttt tggaggtgac caggcctctc
      901 tccaagtggc tgaagcaccc tggggccctg gagaagcaga tgtccagggt agctggagag
      961 tgctggccgc ggccccccac accgcctgcc accaatgtgc tccttatgct ctactccaac
     1021 ctctcgcagg agcagaggca gctgggtggg tccaccttgc tgtgggaagc cgagagctcc
     1081 tggcgggccc aggagggaca gctgtcctgg gagtggggca agaggcaccg gcgacatgag
     1141 cagaaactca tctcagaaga ggatctgcat cacttgccag acagaagtca actgtgtcgg
     1201 aaggtcaagt tccaggtgga cttcaacctg atcggatggg gctcctggat catctacccc
     1261 aagcagtaca acgcctatcg ctgtgagggc gagtgtccta atcctgttgg ggaggagttt
     1321 catccgacca accatgcata catccaggtg gccctgccct gctgtccaag gtcatatggg
     1381 accaaaatgt tttcatttta ctccatgaag tctggaatga gaatttcttg gacttgcaat
     1441 atatcctcca tgccaagcct cagagtctgc tgaacgcgtc atatggctag cctgcaggga
     1501 tccaatgtaa ctgtattcag cgatgacgaa attcttagct attgtaatac tctagaggat
     1561 ctttgtgaag gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat
     1621 ttaaagctct aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct
     1681 aattgtttgt gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa
     1741 tgcctttaat gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc
     1801 tactgctgac tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa
     1861 ggactttcct tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct
     1921 tgcttgcttt gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat
     1981 ggaaaaatat tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt
     2041 ttttcttact ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg
     2101 tacctttagc tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt
     2161 gactagagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2221 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2281 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2341 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     2401 tctgcggcct agtcaataat caatgtcgag acccatagag cccaccgcat ccccagcatg
     2461 cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat
     2521 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt
     2581 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
     2641 gcaactagaa ggcacagtcg aggctgatca gcgggtttaa acgggccctc tagatcaggc
     2701 accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga cgtcggcggt
     2761 gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa
     2821 ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg cgctgcccag
     2881 acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg cgggctcctt
     2941 gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc gggaaccgct
     3001 caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga cgctctccgg
     3061 cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag
     3121 cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg
     3181 atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc
     3241 cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg tcatctcgag
     3301 cctagggccg ggattctcct ccacgtcacc gcatgttaga agacttcctc tgccctctcc
     3361 gctgccagat ctctcgaggc cctgtgggag gaagagaaga ggtcagaagc ttgccccact
     3421 gtggggtgga ggggacagat aaaagtaccc agaaccagag ccacattaac cggccctggg
     3481 aatataaggt ggtcccagct cggggacaca ggatccctgg aggcagcaaa catgctgtcc
     3541 tgaagtggac ataggggccc gggttggagg aagaagacta gctgagctct cggacccctg
     3601 gaagatgcca tgacaggggg ctggaagagc tagcacagac tagagaggta aggggggtag
     3661 gggagctgcc caaatgaaag gagtgagagg tgacccgaat ccacaggaga acggggtgtc
     3721 caggcaaaga aagcaagagg atggagaggt ggctaaagcc agggagacgg ggtactttgg
     3781 ggttgtccag aaaaacggtg atgatgcagg cctacaagaa ggggaggcgg gacgcaaggg
     3841 agacatccgt cggagaaggc catcctaaga aacgagagat ggcacaggcc ccagaaggag
     3901 aaggaaaagg gaacccagcg agtgaagacg gcatggggtt gggtgaggga ggagagatgc
     3961 ccggagagga cccagacacg gggaggatcc gctcagagga catcacgtgg tgcagcgccg
     4021 agaaggaagt gctccggaaa gagcatcctt gggcagcaac acagcagaga gcaaggggaa
     4081 gagggagtgg aggaagacgg aacctgaagg aggcggcagg gaaggatctg ggccagccgt
     4141 agaggtgacc caggccacaa gctgcagaca gaaagcggca caggcccagg ggagagaatg
     4201 ctggtcagag aaagcaaagg gcgaattcgt ttaaacctgc aggactagct agtcccttta
     4261 gtgagggtta attctgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4321 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4381 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4441 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4501 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4561 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4621 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4681 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4741 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4801 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4861 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4921 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4981 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5041 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5101 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5161 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5221 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5281 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5341 ttaagggatt ttggtcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5401 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5461 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5521 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5581 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5641 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5701 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5761 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5821 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5881 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5941 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6001 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6061 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6121 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6181 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6241 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6301 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6421 cacatttccc cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga
     6481 gaaaataccg catcaggaaa ttgtaagcgt taataattca gaagaactcg tcaagaaggc
     6541 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     6601 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     6661 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     6721 ccatgatatt cggcaagcag gcatcgccat gcatgggtca cgacgagatc ctcgccgtcg
     6781 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     6841 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     6901 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     6961 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     7021 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     7081 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt
     7141 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     7201 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     7261 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     7321 aacgatcctc atcctgtctc ttgatcagag cttgatcccc tgcgccatca gatccttggc
     7381 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     7441 gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta
     7501 agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc
     7561 cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtgaaa
     7621 aggatctagg tgaagatcct ttttgataat ctcatgcctg acatttatat tccccagaac
     7681 atcaggttaa tggcgttttt gatgtcattt tcgcggtggc tgagatcagc cacttcttcc
     7741 ccgataacgg agaccggcac actggccata tcggtggtca tcatgcgcca gctttcatcc
     7801 ccgatatgca ccaccgggta aagttcacgg gagactttat ctgacagcag acgtgcactg
     7861 gccaggggga tcaccatccg tcgccccggc gtgtcaataa tatcactctg tacatccaca
     7921 aacagacgat aacggctctc tcttttatag gtgtaaacct taaactgccg tacgtatagg
     7981 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     8041 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     8101 cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga attgaattta
     8161 gcggccgcga attcgccctt agagcagagc caggaacccc tgtagggaag gggcaggaga
     8221 gccaggggca tgagatggtg gacgaggaag ggggacaggg aagcctgagc gcctctcctg
     8281 ggcttgccaa ggactcaaac ccagaagccc agagcagggc cttagggaag cgggaccctg
     8341 ctctgggcgg aggaatatgt cccagatagc actggggact ctttaaggaa agaaggatgg
     8401 agaaagagaa agggagtaga ggcggccacg acctggtgaa cacctaggac gcaccattct
     8461 cacaaaggga gttttccaca cggacacccc cctcctcacc acagccctgc caggacgggg
     8521 ctggctactg gccttatctc acaggtaaaa ctgacgcacg gaggaacaat ataaattggg
     8581 gactagaaag gtgaagagcc aaagttagaa ctcaggacca acttattctg attttgtttt
     8641 tccaaactgc ttctcctctt gggaagtgta aggaagctgc agcaccagga tcagtgaaac
     8701 gcaccagaca gccgcgtcag agcagctcag gttctgggag agggtagcgc agggtggcca
     8761 ctgagaaccg ggcaggtcac gcatcccccc cttccctccc accccctgcc aagctctccc
     8821 tcccaggatc ctctctggct ccatcgtaag caaaccttag aggttctggc aaggagagag
     8881 atggctccag gaaatggggg tgtgtcacca gataaggaat ctgcctaaca ggaggtgggg
     8941 gttagaccca atatcaggag actaggaagg aggaggccta aggatggggc ttttctgtca
     9001 ccaatcctgt ccctagtaaa gcttagtact gtcgatccgc tggcacgaca ggtttcccga
     9061 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     9121 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     9181 atttcacaca ggaaacagct atgaccatga ttacgccaag cttgggctgc aggtcgaggg
     9241 atctccataa gagaagaggg acagctatga ctgggagtag tcaggagagg aggaaaaatc
     9301 tggctagtaa aacatgtaag gaaaatttta gggatgttaa agaaaaaaat aacacaaaac
     9361 aaaatataaa aaaaatctaa cctcaagtca aggcttttct atggaataag gaatggacag
     9421 cagggggctg tttcatatac tgatgacctc tttatagcca acctttgttc atggcagcca
     9481 gcatatgggc atatgttgcc aaactctaaa ccaaatactc attctgatgt tttaaatgat
     9541 ttgccctccc atatgtcctt ccgagtgaga gacacaaaaa attccaacac actattgcaa
     9601 tgaaaataaa tttcctttat tagccagaag tcagatgctc aaggggcttc atgatgtccc
     9661 cataattttt ggcagaggga aaaagatctc agtggtattt gtgagccagg gcattggcca
     9721 caccagccac caccttctga taggcagcct gcacctgagg agtgaattca cgcgccttac
     9781 ccggggagca tgtcaaggtc aaaatcgtca agagcgtcag caggcagcat atcaaggtca
     9841 aagtcgtcaa gggcatcggc tgggagcatg tctaagtcaa aatcgtcaag ggcgtcggtc
     9901 ggcccgccgc tttcgcactt tagctgtttc tccaggccac atatgattag ttccaggccg
     9961 aaaaggaagg caggttcggc tccctgccgg tcgaacagct caattgcttg tttcagaagt
    10021 gggggcatag aatcggtggt aggtgtctct ctttcctctt ttgctacttg atgctcctgt
    10081 tcctccaata cgcagcccag tgtaaagtgg cccacggcgg acagagcgta cagtgcgttc
    10141 tccagggaga agccttgctg acacaggaac gcgagctgat tttccagggt ttcgtactgt
    10201 ttctctgttg ggcgggtgcc gagatgcact ttagccccgt cgcgatgtga gaggagagca
    10261 cagcggtatg acttggcgtt gttccgcaga aagtcttgcc atgactcgcc ttccaggggg
    10321 caggagtggg tatgatgcct gtccagcatc tcgattggca gggcatcgag cagggcccgc
    10381 ttgttcttca cgtgccagta cagggtaggc tgctcaactc ccagcttttg agcgagtttc
    10441 cttgtcgtca ggccttcgat accgactcca ttgagtaatt ccagagcaga gtttatgact
    10501 ttgctcttgt ccagtctaga catggtgctc gacgaattct ttgccaaaat gatgagacag
    10561 cacaataacc agcacgttgc ccaggagctg taggaaaaag aagaaggcat gaacatggtt
    10621 agcagaggct ctagagccgc cggtcacacg ccagaagccg aaccccgccc tgccccgtcc
    10681 cccccgaagg cagccgtccc cccgcggaca gccccgaggc tggagaggga gaaggggacg
    10741 gcggcgcggc gacgcacgaa ggccctcccc gcccatttcc ttcctgccgg cgccgcaccg
    10801 cttcgccccg cgcccgctag agggggtgcg gcggcgcctc ccagatttcg gctccgcaca
    10861 gatttgggac aaaggaagtc cctgcgccct ctcgcacgat taccataaaa ggcaatggct
    10921 gcggctcgcc gcgcctcgac agccgccggc gctccggggg ccgccgcgcc cctcccccga
    10981 gcccyccccg gcccgaggcg gccccgcccc gcccggcacc cccacctgcc gccacccccc
    11041 gcccggcacg gcgagccccg cgccacgccc cgtacggagc cccgcacccg aagccgggcc
    11101 gtgctcagca actcggggag gggggtgcag ggggggttgc agcccgaccg acgcgcccac
    11161 accccctgct caccccccca cgcacacacc ccgcacgcag cctttgttcc cctcgcagcc
    11221 ccccccgcac cgcggggcac cgcccccggc cgcgctcccc tcgcgcacac tgcggagcgc
    11281 acaaagcccc gcgccgcgcc cgcagcgctc acagccgccg ggcagcgcgg agccgcacgc
    11341 ggcgctcccc acgcacacac acacgcacgc accccccgag ccgctccccc cgcacaaagg
    11401 gccctcccgg agcccctcaa ggctttcacg cagccacaga aaagaaacaa gccgtcatta
    11461 aaccaagcgc taattacagc ccggaggaga agggccgtcc cgcccgctca cctgtgggag
    11521 taacgcggtc agtcagagcc ggggcgggcg gcgcgaggcg gcggcggagc ggggcacggg
    11581 gcgaaggcag cgcgcagcga ctcccgcccg ccgcgcgctt cgctttttat agggccgccg
    11641 ccgccgccgc ctcgccataa aaggaaactt tcggagcgcg ccgctctgat tggctgccgc
    11701 cgcacctcyc csccycsccc csccccsccc cycsccccsc cccsccccsc cyggcgcgcg
    11761 cccccccccc ccccgccccc atcgctgcac aaaataatta aaaaataaat aaatacaaaa
    11821 ttgggggtgg ggaggggggg gagatgggga gagtgaagca gaacgtgggg ctcacctcga
    11881 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
    11941 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
    12001 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
    12061 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
    12121 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
    12181 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tacta
//
