LOCUS       sequence-455989-       12379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAVS1 targeting donor plasmid with Flpe-ERT2 expression cassette and
            blasticidin selection gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12379)
  AUTHORS   Chen Y, Cao J, Xiong M, Petersen AJ, Dong Y, Tao Y, Huang CT, Du Z, 
            Zhang SC
  TITLE     Engineering Human Stem Cell Lines with Inducible Gene Knockout using
            CRISPR/Cas9.
  JOURNAL   Cell Stem Cell. 2015 Jul 1. pii: S1934-5909(15)00261-1. doi: 
            10.1016/j.stem.2015.06.001.
  PUBMED    26145478
REFERENCE   2  (bases 1 to 12379)
  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: "Cell Stem 
            Cell. 2015 Jul 1. pii: S1934-5909(15)00261-1. doi: 
            10.1016/j.stem.2015.06.001."
COMMENT     Sequence Label: AAVS1-Blasticidin-CAG-Flpe-ERT2
FEATURES             Location/Qualifiers
     source          1..12379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    33..836
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    843..868
                     /label=SA
                     /note="splice acceptor site"
     CDS             892..945
                     /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"
     primer_bind     complement(1418..1435)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1424..1648
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1716..2095
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2097..2375
                     /label=chicken beta-actin promoter
     intron          2376..3392
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3316..3338
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3400..3419
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3456..3465
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4761..5696
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             4809..5702
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     misc_feature    5736..6324
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5789..5809)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6207..6218)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6356..6832
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6555..6574)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6942..6961)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7007..7062
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7061..7080)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7423..7439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7447..7463
                     /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(7471..7477,7478..7495,7496..7501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7516..7537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    7611..8447
                     /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(8481..8500)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8482..8500)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8507..8524)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8507..8523)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8516..8538)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8661..8963
                     /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     8842..8861
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(9366..9385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9980..9999
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(10360..11151,11152..11220))
                     /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     10983..11002
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11344..11932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11833..11852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12086..12103
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12220..12241
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12256..12261,12262..12279,12280..12286)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12294..12310
                     /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     12318..12334
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        12355..12373
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctagtcctgc aggtttaaac gaattcgccc tttgctttct ctgaccagca ttctctcccc
       61 tgggcctgtg ccgctttctg tctgcagctt gtggcctggg tcacctctac ggctggccca
      121 gatccttccc tgccgcctcc ttcaggttcc gtcttcctcc actccctctt ccccttgctc
      181 tctgctgtgt tgctgcccaa ggatgctctt tccggagcac ttccttctcg gcgctgcacc
      241 acgtgatgtc ctctgagcgg atcctccccg tgtctgggtc ctctccgggc atctctcctc
      301 cctcacccaa ccccatgccg tcttcactcg ctgggttccc ttttccttct ccttctgggg
      361 cctgtgccat ctctcgtttc ttaggatggc cttctccgac ggatgtctcc cttgcgtccc
      421 gcctcccctt cttgtaggcc tgcatcatca ccgtttttct ggacaacccc aaagtacccc
      481 gtctccctgg ctttagccac ctctccatcc tcttgctttc tttgcctgga caccccgttc
      541 tcctgtggat tcgggtcacc tctcactcct ttcatttggg cagctcccct acccccctta
      601 cctctctagt ctgtgctagc tcttccagcc ccctgtcatg gcatcttcca ggggtccgag
      661 agctcagcta gtcttcttcc tccaacccgg gcccctatgt ccacttcagg acagcatgtt
      721 tgctgcctcc agggatcctg tgtccccgag ctgggaccac cttatattcc cagggccggt
      781 taatgtggct ctggttctgg gtacttttat ctgtcccctc caccccacag tggggcaagc
      841 ttctgacctc ttctcttcct cccacagggc ctcgagagat ctggcagcgg agagggcaga
      901 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctaggac cggtatgaaa
      961 acatttaaca tttctcaaca agatctagaa ttagtagaag tagcgacaga gaagattaca
     1021 atgctttatg aggataataa acatcatgtg ggagcggcaa ttcgtacgaa aacaggagaa
     1081 atcatttcgg cagtacatat tgaagcgtat ataggacgag taactgtttg tgcagaagcc
     1141 attgcgattg gtagtgcagt ttcgaatgga caaaaggatt ttgacacgat tgtagctgtt
     1201 agacaccctt attctgacga agtagataga agtattcgag tggtaagtcc ttgtggtatg
     1261 tgtagggagt tgatttcaga ctatgcacca gattgttttg tgttaataga aatgaatggc
     1321 aagttagtca aaactacgat tgaagaactc attccactca aatatacccg aaattaatga
     1381 gaattcgggg atcaattctc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     1441 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     1501 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     1561 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     1621 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg
     1681 actagagctt gcggaaccct taatggtacc gtctcgacat tgattattga ctagttatta
     1741 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     1801 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     1861 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     1921 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     1981 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     2041 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
     2101 ggtgagcccc acgttctgct tcactctccc catctccccc cccctcccca cccccaattt
     2161 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc
     2221 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
     2281 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
     2341 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
     2401 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     2461 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
     2521 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
     2581 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
     2641 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
     2701 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     2761 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     2821 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     2881 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     2941 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     3001 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     3061 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     3121 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     3181 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     3241 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     3301 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     3361 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     3421 tgtctcatca ttttggcaaa gaattcgtcg acgccgccac catggctatg agccagttcg
     3481 acatcctgtg caagaccccc cccaaggtgc tggtgcggca gttcgtggag agattcgaga
     3541 ggcccagcgg cgagaagatc gccagctgtg ccgccgagct gacctacctg tgctggatga
     3601 tcacccacaa cggcaccgcc atcaagaggg ccaccttcat gagctacaac accatcatca
     3661 gcaacagcct gagcttcgac atcgtgaaca agagcctgca gttcaagtac aagacccaga
     3721 aggccaccat cctggaggcc agcctgaaga agctgatccc cgcctgggag ttcaccatca
     3781 tcccttacaa cggccagaag caccagagcg acatcaccga catcgtgtcc agcctgcagc
     3841 tgcagttcga gagcagcgag gaggccgaca agggcaacag ccacagcaag aagatgctga
     3901 aggccctgct gtccgagggc gagagcatct gggagatcac cgagaagatc ctgaacagct
     3961 tcgagtacac cagcaggttc accaagacca agaccctgta ccagttcctg ttcctggcca
     4021 cattcatcaa ctgcggcagg ttcagcgaca tcaagaacgt ggaccccaag agcttcaagc
     4081 tggtgcagaa caagtacctg ggcgtgatca ttcagtgcct ggtgaccgag accaagacaa
     4141 gcgtgtccag gcacatctac tttttcagcg ccagaggcag gatcgacccc ctggtgtacc
     4201 tggacgagtt cctgaggaac agcgagcccg tgctgaagag agtgaacagg accggcaaca
     4261 gcagcagcaa caagcaggag taccagctgc tgaaggacaa cctggtgcgc agctacaaca
     4321 aggccctgaa gaagaacgcc ccctacccca tcttcgctat caagaacggc cctaagagcc
     4381 acatcggcag gcacctgatg accagctttc tgagcatgaa gggcctgacc gagctgacaa
     4441 acgtggtggg caactggagc gacaagaggg cctccgccgt ggccaggacc acctacaccc
     4501 accagatcac cgccatcccc gaccactact tcgccctggt gtccaggtac tacgcctacg
     4561 accccatcag caaggagatg atcgccctga aggacgagac caaccccatc gaggagtggc
     4621 agcacatcga gcagctgaag ggcagcgccg agggcagcat cagatacccc gcctggaacg
     4681 gcatcatcag ccaggaggtg ctggactacc tgagcagcta catcaacagg cggatcaccg
     4741 gttgcgtacg cggatcctct gctggagaca tgagagctgc caacctttgg ccaagcccgc
     4801 tcatgatcaa acgctctaag aagaacagcc tggccttgtc cctgacggcc gaccagatgg
     4861 tcagtgcctt gttggatgct gagcccccca tactctattc cgagtatgat cctaccagac
     4921 ccttcagtga agcttcgatg atgggcttac tgaccaacct ggcagacagg gagctggttc
     4981 acatgatcaa ctgggcgaag agggtgccag gctttgtgga tttgaccctc catgatcagg
     5041 tccaccttct agaatgtgcc tggctagaga tcctgatgat tggtctcgtc tggcgctcca
     5101 tggagcaccc agtgaagcta ctgtttgctc ctaacttgct cttggacagg aaccagggaa
     5161 aatgtgtaga gggcatggtg gagatcttcg acatgctgct ggctacatca tctcggttcc
     5221 gcatgatgaa tctgcaggga gaggagtttg tgtgcctcaa atctattatt ttgcttaatt
     5281 ctggagtgta cacatttctg tccagcaccc tgaagtctct ggaagagaag gaccatatcc
     5341 accgagtcct ggacaagatc acagacactt tgatccacct gatggccaag gcaggcctga
     5401 ccctgcagca gcagcaccag cggctggccc agctcctcct catcctctcc cacatcaggc
     5461 acatgagtaa caaaggcatg gagcatctgt acagcatgaa gtgcaagaac gtggtgcccc
     5521 tctatgacct gctgctggag gcggcggacg cccaccgcct acatgcgccc actagccgtg
     5581 gaggggcatc cgtggaggag acggaccaaa gccacttggc cactgcgggc tctacttcat
     5641 cgcattcctt gcaaaagtat tacatcacgg gggaggcaga gggtttccct gccacagctt
     5701 gatagacgcg tgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     5761 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     5821 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     5881 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     5941 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     6001 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     6061 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     6121 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     6181 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     6241 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     6301 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     6361 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     6421 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     6481 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     6541 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     6601 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     6661 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     6721 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     6781 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     6841 gtaggtaacc aggcgcgtga attcactcct caggtgcagg ctgcctatca gaaggtggtg
     6901 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
     6961 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     7021 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     7081 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     7141 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     7201 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     7261 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     7321 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     7381 tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
     7441 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     7501 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7561 tttccagtcg ggaaacctgt cgtgccagcg gatcgacagt actaagcttt actagggaca
     7621 ggattggtga cagaaaagcc ccatccttag gcctcctcct tcctagtctc ctgatattgg
     7681 gtctaacccc cacctcctgt taggcagatt ccttatctgg tgacacaccc ccatttcctg
     7741 gagccatctc tctccttgcc agaacctcta aggtttgctt acgatggagc cagagaggat
     7801 cctgggaggg agagcttggc agggggtggg agggaagggg gggatgcgtg acctgcccgg
     7861 ttctcagtgg ccaccctgcg ctaccctctc ccagaacctg agctgctctg acgcggctgt
     7921 ctggtgcgtt tcactgatcc tggtgctgca gcttccttac acttcccaag aggagaagca
     7981 gtttggaaaa acaaaatcag aataagttgg tcctgagttc taactttggc tcttcacctt
     8041 tctagtcccc aatttatatt gttcctccgt gcgtcagttt tacctgtgag ataaggccag
     8101 tagccagccc cgtcctggca gggctgtggt gaggaggggg gtgtccgtgt ggaaaactcc
     8161 ctttgtgaga atggtgcgtc ctaggtgttc accaggtcgt ggccgcctct actccctttc
     8221 tctttctcca tccttctttc cttaaagagt ccccagtgct atctgggaca tattcctccg
     8281 cccagagcag ggtcccgctt ccctaaggcc ctgctctggg cttctgggtt tgagtccttg
     8341 gcaagcccag gagaggcgct caggcttccc tgtccccctt cctcgtccac catctcatgc
     8401 ccctggctct cctgcccctt ccctacaggg gttcctggct ctgctctaag ggcgaattcg
     8461 cggccgctaa attcaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt
     8521 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     8581 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     8641 tgcgcagcct atacgtacgg cagtttaagg tttacaccta taaaagagag agccgttatc
     8701 gtctgtttgt ggatgtacag agtgatatta ttgacacgcc ggggcgacgg atggtgatcc
     8761 ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc
     8821 atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg
     8881 ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta
     8941 acctgatgtt ctggggaata taaatgtcag gcatgagatt atcaaaaagg atcttcacct
     9001 agatcctttt cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca
     9061 gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca
     9121 gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat
     9181 tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt
     9241 tcttgccgcc aaggatctga tggcgcaggg gatcaagctc tgatcaagag acaggatgag
     9301 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     9361 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     9421 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     9481 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     9541 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     9601 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     9661 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     9721 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     9781 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     9841 gcatgcccga cggcgaggat ctcgtcgtga cccatgcatg gcgatgcctg cttgccgaat
     9901 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     9961 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
    10021 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
    10081 ttctatcgcc ttcttgacga gttcttctga attattaacg cttacaattt cctgatgcgg
    10141 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tcaggtggca cttttcgggg
    10201 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
    10261 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
    10321 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
    10381 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
    10441 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
    10501 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
    10561 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
    10621 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
    10681 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
    10741 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
    10801 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    10861 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg aatactcaac
    10921 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    10981 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    11041 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    11101 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    11161 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    11221 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgaccaaaat
    11281 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
    11341 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
    11401 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
    11461 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
    11521 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
    11581 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
    11641 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
    11701 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
    11761 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
    11821 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
    11881 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
    11941 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
    12001 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
    12061 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
    12121 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
    12181 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
    12241 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
    12301 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tcagaattaa
    12361 ccctcactaa agggactag
//
