LOCUS       sequence-264416-       11878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  donor plasmid for cardiac-specific FUCCI expression in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11878)
  AUTHORS   Chang Y, Hellwarth PB, Randolph LN, Sun Y, Xing Y, Zhu W, Lian XL, 
            Bao X
  TITLE     Fluorescent indicators for continuous and lineage-specific reporting
            of cell-cycle phases in human pluripotent stem cells.
  JOURNAL   Biotechnol Bioeng. 2020 Apr 11. doi: 10.1002/bit.27352.
  PUBMED    32277708
REFERENCE   2  (bases 1 to 11878)
  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: "Biotechnol 
            Bioeng. 2020 Apr 11. doi: 10.1002/bit.27352."
COMMENT     Sequence Label: AAVS1-Puro cTnT FUCCI
FEATURES             Location/Qualifiers
     source          1..11878
                     /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"
     CDS             955..1554
                     /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(955..974)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1451..1471
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1586..1603)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1592..1816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2460..2478
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2510..2534
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(2550..2552,2553..2555,2556..3266)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2595..2616)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3203..3224
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3297..3626
                     /codon_start=1
                     /product="degron consisting of residues 1-110 of human 
                     Geminin (Zielke and Edgar, 2015)"
                     /label=Gem1 (1-110)
                     /translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS
                     AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE
                     KRRKAL"
     misc_feature    3631..4203
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3798..3815)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4025..4044
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4218..4868
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     CDS             4218..4868
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     CDS             4923..5195
                     /codon_start=1
                     /product="degron consisting of residues 30-120 of human 
                     Cdt1 (Zielke and Edgar, 2015)"
                     /label=Cdt1 (30-120)
                     /translation="PSPARPALRAPASATSGSRKRARPPAAPGRDQARPPARRRLRLSV
                     DEVSSPSTPEAPDIPACPSPGQKIKKSTPAAGQPPHLTSAQDQDTI"
     misc_feature    5233..5821
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5286..5306)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5704..5715)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5853..6329
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6052..6071)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6439..6458)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6504..6559
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6558..6577)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6920..6936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6944..6960
                     /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(6968..6974,6975..6992,6993..6998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7013..7034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    7108..7944
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(7985..8003)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8010..8027)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8010..8026)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8019..8041)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8164..8466
                     /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     8345..8364
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(8869..8888)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9483..9502
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(9863..10654,10655..10723))
                     /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     10486..10505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10847..11435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11336..11355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11589..11606
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11723..11744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11759..11764,11765..11782,11783..11789)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11797..11813
                     /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     11821..11837
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11858..11876
                     /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 gccctaggct cgagatgacc
      961 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     1021 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     1081 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     1141 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     1201 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     1261 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     1321 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     1381 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     1441 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     1501 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgatctaga
     1561 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     1621 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1681 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1741 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     1801 gcggtgggct ctatgggtct cgacattgat tattgactag tgtcatggag aagacccacc
     1861 ttgcagatgt cctcactggg gctggcagag ccggcaacct gcctaaggct gctcagtcca
     1921 ttaggagcca gtagcctgga agatgtcttt acccccagca tcagttcaag tggagcagca
     1981 cataactctt gccctctgcc ttccaagatt ctggtgctga gacttatgga gtgtcttgga
     2041 ggttgccttc tgccccccaa ccctgctccc agctggccct cccaggcctg ggttgctggc
     2101 ctctgcttta tcaggattct caagagggac agctggttta tgttgcatga ctgttccctg
     2161 catatctgct ctggttttaa atagcttatc tgagcagctg gaggaccaca tgggcttata
     2221 tggcgtgggg tacatgttcc tgtagccttg tccctggcac ctgccaaaat agcagccaac
     2281 accccccacc cccaccgcca tccccctgcc ccacccgtcc cctgtcgcac attcctccct
     2341 ccgcagggct ggctcaccag gccccagccc acatgcctgc ttaaagccct ctccatcctc
     2401 tgcctcaccc agtccccgct gagactgagc agacgcctcc aggatccgcc accctcgagt
     2461 aatacgactc actataggga gacccaagct ggctaggtaa gcttgatcaa caagtttgta
     2521 caaaaaagca ggctgtcgac gctagcacca tggtgagcaa gggcgaggag ctgttcaccg
     2581 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtcc
     2641 gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca
     2701 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcgtggcct
     2761 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2821 aaggctacgt ccaggagcgc accatctctt tcaaggacga cggtacctac aagacccgcg
     2881 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2941 tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac agccacaacg
     3001 tctatatcac ggccgacaag cagaagaacg gcatcaaggc taacttcaag atccgccaca
     3061 acgttgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3121 acggccccgt gctgctgccc gacaaccact acctgagcca tcagtccgcc ctgagcaaag
     3181 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatta
     3241 cacatggcat ggacgagctg tacaaggata tccatgcaca ctggcggccg ctcgagatga
     3301 atcccagtat gaagcagaaa caagaagaaa tcaaagagaa tataaagaat agttctgtcc
     3361 caagaagaac tctgaagatg attcagcctt ctgcatctgg atctcttgtt ggaagagaaa
     3421 atgagctgtc cgcaggcttg tccaaaagga aacatcggaa tgaccactta acatctacaa
     3481 cttccagccc tggggttatt gtcccagaat ctagtgaaaa taaaaatctt ggaggagtca
     3541 cccaggagtc atttgatctt atgattaaag aaaatccatc ctctcagtat tggaaggaag
     3601 tggcagaaaa acggagaaag gcgctgtaag cccctctccc tccccccccc ctaacgttac
     3661 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     3721 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     3781 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     3841 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     3901 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     3961 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     4021 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     4081 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     4141 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     4201 taatggccac agaattcatg gtgagcgtga tcaagcccga gatgaagatg aggtactaca
     4261 tggacggctc cgtcaatggg catgagttca caatcgaggg tgagggcaca ggcagacctt
     4321 acgagggaca tcaggagatg acactgcgcg tcacaatggc cgagggcggg ccaatgcctt
     4381 tcgccttcga cctggtgtcc cacgtgttct gttacggcca cagagttttt accaagtacc
     4441 cagaagagat cccagactat ttcaagcagg cctttcctga gggcctgtcc tgggagaggt
     4501 ccctggagtt cgaggacggc ggctccgcct ccgtgagcgc ccacatcagc ctgaggggca
     4561 acaccttcta ccacaagtcc aagttcaccg gcgtgaactt ccccgccgac ggccccatca
     4621 tgcagaacca gagcgtggac tgggagccct ccaccgagaa gatcaccgcc agcgacggcg
     4681 tgctgaaggg cgacgtgacc atgtacctga agctggaggg cggcggcaac cacaagtgcc
     4741 agatgaagac cacctacaag gccgccaagg agatcctgga gatgcccggc gaccactaca
     4801 tcggccacag gctggtgagg aagaccgagg gcaacatcac cgagcaggtg gaggacgccg
     4861 tggcccacta ctcaatgttg ccttccctgg gcgatatcca tgcacactgg cggccgctcg
     4921 agcccagccc cgccaggccc gcactccgcg ccccggcctc cgctaccagt ggcagccgca
     4981 agcgcgcccg cccgcccgcc gcccccggac gcgaccaggc caggccaccg gcccgcagga
     5041 gactgcggct gtcggtggac gaggtttcca gccccagtac ccccgaggcc ccagacatcc
     5101 cagcctgccc ttctccgggc cagaagataa agaaatccac cccggcagca ggtcagccgc
     5161 cccacctgac atccgcgcag gaccaggaca ccatctgagt ttacgcgtga attcgatatc
     5221 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5281 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5341 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5401 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5461 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5521 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5581 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     5641 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5701 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5761 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     5821 catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag
     5881 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     5941 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     6001 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     6061 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     6121 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     6181 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     6241 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     6301 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccagg cgcgtgaatt
     6361 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     6421 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     6481 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     6541 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     6601 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     6661 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     6721 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     6781 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     6841 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     6901 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6961 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7021 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7081 gccagcggat cgacagtact aagctttact agggacagga ttggtgacag aaaagcccca
     7141 tccttaggcc tcctccttcc tagtctcctg atattgggtc taacccccac ctcctgttag
     7201 gcagattcct tatctggtga cacaccccca tttcctggag ccatctctct ccttgccaga
     7261 acctctaagg tttgcttacg atggagccag agaggatcct gggagggaga gcttggcagg
     7321 gggtgggagg gaaggggggg atgcgtgacc tgcccggttc tcagtggcca ccctgcgcta
     7381 ccctctccca gaacctgagc tgctctgacg cggctgtctg gtgcgtttca ctgatcctgg
     7441 tgctgcagct tccttacact tcccaagagg agaagcagtt tggaaaaaca aaatcagaat
     7501 aagttggtcc tgagttctaa ctttggctct tcacctttct agtccccaat ttatattgtt
     7561 cctccgtgcg tcagttttac ctgtgagata aggccagtag ccagccccgt cctggcaggg
     7621 ctgtggtgag gaggggggtg tccgtgtgga aaactccctt tgtgagaatg gtgcgtccta
     7681 ggtgttcacc aggtcgtggc cgcctctact ccctttctct ttctccatcc ttctttcctt
     7741 aaagagtccc cagtgctatc tgggacatat tcctccgccc agagcagggt cccgcttccc
     7801 taaggccctg ctctgggctt ctgggtttga gtccttggca agcccaggag aggcgctcag
     7861 gcttccctgt cccccttcct cgtccaccat ctcatgcccc tggctctcct gccccttccc
     7921 tacaggggtt cctggctctg ctctaagggc aagggcgaat tcgcggccgc taaattcaat
     7981 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     8041 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     8101 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctatacgta
     8161 cggcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     8221 cagagtgata ttattgacac gccggggcga cggatggtga tccccctggc cagtgcacgt
     8281 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     8341 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     8401 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     8461 atataaatgt caggcatgag attatcaaaa aggatcttca cctagatcct tttcacgtag
     8521 aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg
     8581 acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga
     8641 tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc
     8701 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
     8761 tgatggcgca ggggatcaag ctctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     8821 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     8881 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     8941 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     9001 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     9061 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     9121 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     9181 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     9241 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     9301 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     9361 gatctcgtcg tgacccatgc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     9421 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     9481 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     9541 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     9601 cgagttcttc tgaattatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     9661 tctgtgcggt atttcacacc gcatcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     9721 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga ttatcaaaaa
     9781 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     9841 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     9901 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     9961 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
    10021 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
    10081 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
    10141 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
    10201 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
    10261 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
    10321 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
    10381 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
    10441 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
    10501 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
    10561 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
    10621 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    10681 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
    10741 attattgaag catttatcag ggttattgtc tcatgaccaa aatcccttaa cgtgagtttt
    10801 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
    10861 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
    10921 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
    10981 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
    11041 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
    11101 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
    11161 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
    11221 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
    11281 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
    11341 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
    11401 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
    11461 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
    11521 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
    11581 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
    11641 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
    11701 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
    11761 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
    11821 caggaaacag ctatgaccat gattacgcca agctcagaat taaccctcac taaagggt
//
