LOCUS       sequence-272078-       11876 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 11876)
  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 11876)
  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..11876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..837
                     /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    911..932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(947..952,953..970,971..977)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    985..1001
                     /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     1009..1025
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1368..1387
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1386..1441
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1487..1506
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1616..2092
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1874..1893
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    2124..2712
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2230..2241
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2639..2659
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2750..3022)
                     /codon_start=1
                     /product="degron consisting of residues 30-120 of human 
                     Cdt1 (Zielke and Edgar, 2015)"
                     /label=Cdt1 (30-120)
                     /translation="PSPARPALRAPASATSGSRKRARPPAAPGRDQARPPARRRLRLSV
                     DEVSSPSTPEAPDIPACPSPGQKIKKSTPAAGQPPHLTSAQDQDTI"
     CDS             complement(3077..3727)
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     CDS             complement(3077..3727)
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     misc_feature    3742..4314
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3901..3920)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     4130..4147
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(4319..4648)
                     /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"
     CDS             complement(join(4679..5389,5390..5392,5393..5395))
                     /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(4721..4742)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5329..5350
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(5411..5435)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(5467..5485)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6129..6353
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6342..6359
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(6391..6990)
                     /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(6474..6494)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6971..6990
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(7000..7053)
                     /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    7077..7102
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    7109..7912
                     /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(7951..7969)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7990..8006)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8014..8030
                     /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(8038..8044,8045..8062,8063..8068))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8083..8104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8221..8238)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8392..8980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8472..8491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(9104..9172,9173..9964)
                     /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(9322..9341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(10325..10344)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     10939..10958
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(11361..11663)
                     /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(11463..11482)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     11786..11808
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11800..11817
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11801..11817
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11824..11842
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agagcagagc caggaacccc tgtagggaag gggcaggaga gccaggggca tgagatggtg
       61 gacgaggaag ggggacaggg aagcctgagc gcctctcctg ggcttgccaa ggactcaaac
      121 ccagaagccc agagcagggc cttagggaag cgggaccctg ctctgggcgg aggaatatgt
      181 cccagatagc actggggact ctttaaggaa agaaggatgg agaaagagaa agggagtaga
      241 ggcggccacg acctggtgaa cacctaggac gcaccattct cacaaaggga gttttccaca
      301 cggacacccc cctcctcacc acagccctgc caggacgggg ctggctactg gccttatctc
      361 acaggtaaaa ctgacgcacg gaggaacaat ataaattggg gactagaaag gtgaagagcc
      421 aaagttagaa ctcaggacca acttattctg attttgtttt tccaaactgc ttctcctctt
      481 gggaagtgta aggaagctgc agcaccagga tcagtgaaac gcaccagaca gccgcgtcag
      541 agcagctcag gttctgggag agggtagcgc agggtggcca ctgagaaccg ggcaggtcac
      601 gcatcccccc cttccctccc accccctgcc aagctctccc tcccaggatc ctctctggct
      661 ccatcgtaag caaaccttag aggttctggc aaggagagag atggctccag gaaatggggg
      721 tgtgtcacca gataaggaat ctgcctaaca ggaggtgggg gttagaccca atatcaggag
      781 actaggaagg aggaggccta aggatggggc ttttctgtca ccaatcctgt ccctagtaaa
      841 gcttagtact gtcgatccgc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
      901 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
      961 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     1021 atgaccatga ttacgccaag cttgggctgc aggtcgaggg atctccataa gagaagaggg
     1081 acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa aacatgtaag
     1141 gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatataaa aaaaatctaa
     1201 cctcaagtca aggcttttct atggaataag gaatggacag cagggggctg tttcatatac
     1261 tgatgacctc tttatagcca acctttgttc atggcagcca gcatatgggc atatgttgcc
     1321 aaactctaaa ccaaatactc attctgatgt tttaaatgat ttgccctccc atatgtcctt
     1381 ccgagtgaga gacacaaaaa attccaacac actattgcaa tgaaaataaa tttcctttat
     1441 tagccagaag tcagatgctc aaggggcttc atgatgtccc cataattttt ggcagaggga
     1501 aaaagatctc agtggtattt gtgagccagg gcattggcca caccagccac caccttctga
     1561 taggcagcct gcacctgagg agtgaattca cgcgcctggt tacctacaaa atcagaagga
     1621 cagggaaggg agcagtggtt cacgcctgta atcccagcaa tttgggaggc caaggtgggt
     1681 agatcacctg agattaggag ttggagacca gcctggccaa tatggtgaaa ccccgtctct
     1741 accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc ctggaatccc aacaactcgg
     1801 gaggctgagg caggagaatc gcttgaaccc aggaggcgga gattgcagtg agccaagatt
     1861 gtgccactgc actccagctt ggttcccaat agaccccgca ggccctacag gttgtcttcc
     1921 caacttgccc cttgctccat accacccccc tccaccccat aatattatag aaggacacct
     1981 agtcagacaa aatgatgcaa cttaatttta ttaggacaag gctggtgggc actggagtgg
     2041 caacttccag ggccaggaga ggcactgggg aggggtcaca gggatgccac ccgtagatct
     2101 ctcgagcagc gctcggtatc gatgcgggga ggcggcccaa agggagatcc gactcgtctg
     2161 agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag
     2221 gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt
     2281 ggcaacatag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca ccacggaatt
     2341 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
     2401 cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac aggtggtggc
     2461 aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct
     2521 gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa
     2581 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
     2641 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
     2701 cagaggttga ttatcgataa gcttgatatc gaattcacgc gtaaactcag atggtgtcct
     2761 ggtcctgcgc ggatgtcagg tggggcggct gacctgctgc cggggtggat ttctttatct
     2821 tctggcccgg agaagggcag gctgggatgt ctggggcctc gggggtactg gggctggaaa
     2881 cctcgtccac cgacagccgc agtctcctgc gggccggtgg cctggcctgg tcgcgtccgg
     2941 gggcggcggg cgggcgggcg cgcttgcggc tgccactggt agcggaggcc ggggcgcgga
     3001 gtgcgggcct ggcggggctg ggctcgagcg gccgccagtg tgcatggata tcgcccaggg
     3061 aaggcaacat tgagtagtgg gccacggcgt cctccacctg ctcggtgatg ttgccctcgg
     3121 tcttcctcac cagcctgtgg ccgatgtagt ggtcgccggg catctccagg atctccttgg
     3181 cggccttgta ggtggtcttc atctggcact tgtggttgcc gccgccctcc agcttcaggt
     3241 acatggtcac gtcgcccttc agcacgccgt cgctggcggt gatcttctcg gtggagggct
     3301 cccagtccac gctctggttc tgcatgatgg ggccgtcggc ggggaagttc acgccggtga
     3361 acttggactt gtggtagaag gtgttgcccc tcaggctgat gtgggcgctc acggaggcgg
     3421 agccgccgtc ctcgaactcc agggacctct cccaggacag gccctcagga aaggcctgct
     3481 tgaaatagtc tgggatctct tctgggtact tggtaaaaac tctgtggccg taacagaaca
     3541 cgtgggacac caggtcgaag gcgaaaggca ttggcccgcc ctcggccatt gtgacgcgca
     3601 gtgtcatctc ctgatgtccc tcgtaaggtc tgcctgtgcc ctcaccctcg attgtgaact
     3661 catgcccatt gacggagccg tccatgtagt acctcatctt catctcgggc ttgatcacgc
     3721 tcaccatgaa ttctgtggcc attatcatcg tgtttttcaa aggaaaacca cgtccccgtg
     3781 gttcgggggg cctagacgtt ttttaacctc gactaaacac atgtaaagca tgtgcaccga
     3841 ggccccagat cagatcccat acaatggggt accttctggg catccttcag ccccttgttg
     3901 aatacgcttg aggagagcca tttgactctt tccacaacta tccaactcac aacgtggcac
     3961 tggggttgtg ccgcctttgc aggtgtatct tatacacgtg gcttttggcc gcagaggcac
     4021 ctgtcgccag gtggggggtt ccgctgcctg caaagggtcg ctacagacgt tgtttgtctt
     4081 caagaagctt ccagaggaac tgcttccttc acgacattca acagaccttg cattcctttg
     4141 gcgagagggg aaagacccct aggaatgctc gtcaagaaga cagggccagg tttccgggcc
     4201 ctcacattgc caaaagacgg caatatggtg gaaaataaca tatagacaaa cgcacaccgg
     4261 ccttattcca agcggcttcg gccagtaacg ttaggggggg gggagggaga ggggcttaca
     4321 gcgcctttct ccgtttttct gccacttcct tccaatactg agaggatgga ttttctttaa
     4381 tcataagatc aaatgactcc tgggtgactc ctccaagatt tttattttca ctagattctg
     4441 ggacaataac cccagggctg gaagttgtag atgttaagtg gtcattccga tgtttccttt
     4501 tggacaagcc tgcggacagc tcattttctc ttccaacaag agatccagat gcagaaggct
     4561 gaatcatctt cagagttctt cttgggacag aactattctt tatattctct ttgatttctt
     4621 cttgtttctg cttcatactg ggattcatct cgagcggccg ccagtgtgca tggatatcct
     4681 tgtacagctc gtccatgcca tgtgtaatcc cggcggcggt cacgaactcc agcaggacca
     4741 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgatggctc aggtagtggt
     4801 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     4861 gctgcacgct gccgtcctca acgttgtggc ggatcttgaa gttagccttg atgccgttct
     4921 tctgcttgtc ggccgtgata tagacgttgt ggctgttgaa gttgtactcc agcttgtgcc
     4981 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     5041 tgtcgccctc gaacttcacc tcggcgcggg tcttgtaggt accgtcgtcc ttgaaagaga
     5101 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     5161 tgtggtcggg gtagcggctg aagcaggcca cgccgtagcc gaaggtggtc acgagggtgg
     5221 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgttgg
     5281 tggcatcgcc ctcgccctcg ccgcggacgc tgaacttgtg gccgtttacg tcgccgtcca
     5341 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgc
     5401 tagcgtcgac agcctgcttt tttgtacaaa cttgttgatc aagcttacct agccagcttg
     5461 ggtctcccta tagtgagtcg tattactcga gggtggcgga tcctggaggc gtctgctcag
     5521 tctcagcggg gactgggtga ggcagaggat ggagagggct ttaagcaggc atgtgggctg
     5581 gggcctggtg agccagccct gcggagggag gaatgtgcga caggggacgg gtggggcagg
     5641 gggatggcgg tgggggtggg gggtgttggc tgctattttg gcaggtgcca gggacaaggc
     5701 tacaggaaca tgtaccccac gccatataag cccatgtggt cctccagctg ctcagataag
     5761 ctatttaaaa ccagagcaga tatgcaggga acagtcatgc aacataaacc agctgtccct
     5821 cttgagaatc ctgataaagc agaggccagc aacccaggcc tgggagggcc agctgggagc
     5881 agggttgggg ggcagaaggc aacctccaag acactccata agtctcagca ccagaatctt
     5941 ggaaggcaga gggcaagagt tatgtgctgc tccacttgaa ctgatgctgg gggtaaagac
     6001 atcttccagg ctactggctc ctaatggact gagcagcctt gggcaggttg ccggctctgc
     6061 cagccccagt gaggacatct gcaaggtggg tcttctccat gacactagtc aataatcaat
     6121 gtcgagaccc atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc
     6181 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
     6241 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
     6301 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc
     6361 tgatcagcgg gtttaaacgg gccctctaga tcaggcaccg ggcttgcggg tcatgcacca
     6421 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
     6481 gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc
     6541 cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
     6601 gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
     6661 ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
     6721 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc
     6781 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc
     6841 ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg
     6901 gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc
     6961 gaggcgcacc gtgggcttgt actcggtcat ctcgagccta gggccgggat tctcctccac
     7021 gtcaccgcat gttagaagac ttcctctgcc ctctccgctg ccagatctct cgaggccctg
     7081 tgggaggaag agaagaggtc agaagcttgc cccactgtgg ggtggagggg acagataaaa
     7141 gtacccagaa ccagagccac attaaccggc cctgggaata taaggtggtc ccagctcggg
     7201 gacacaggat ccctggaggc agcaaacatg ctgtcctgaa gtggacatag gggcccgggt
     7261 tggaggaaga agactagctg agctctcgga cccctggaag atgccatgac agggggctgg
     7321 aagagctagc acagactaga gaggtaaggg gggtagggga gctgcccaaa tgaaaggagt
     7381 gagaggtgac ccgaatccac aggagaacgg ggtgtccagg caaagaaagc aagaggatgg
     7441 agaggtggct aaagccaggg agacggggta ctttggggtt gtccagaaaa acggtgatga
     7501 tgcaggccta caagaagggg aggcgggacg caagggagac atccgtcgga gaaggccatc
     7561 ctaagaaacg agagatggca caggccccag aaggagaagg aaaagggaac ccagcgagtg
     7621 aagacggcat ggggttgggt gagggaggag agatgcccgg agaggaccca gacacgggga
     7681 ggatccgctc agaggacatc acgtggtgca gcgccgagaa ggaagtgctc cggaaagagc
     7741 atccttgggc agcaacacag cagagagcaa ggggaagagg gagtggagga agacggaacc
     7801 tgaaggaggc ggcagggaag gatctgggcc agccgtagag gtgacccagg ccacaagctg
     7861 cagacagaaa gcggcacagg cccaggggag agaatgctgg tcagagaaag caaagggcga
     7921 attcgtttaa acctgcagga ctagctagtc cctttagtga gggttaattc tgagcttggc
     7981 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     8041 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     8101 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     8161 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     8221 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     8281 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     8341 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     8401 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     8461 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     8521 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     8581 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     8641 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     8701 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     8761 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8821 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     8881 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8941 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     9001 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagaca
     9061 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     9121 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     9181 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     9241 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     9301 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     9361 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     9421 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     9481 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     9541 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     9601 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     9661 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     9721 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     9781 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     9841 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     9901 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     9961 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
    10021 atttaaaagg atctaggtga agatcctttt tgataatctc atgagcggat acatatttga
    10081 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    10141 tgatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
    10201 aagcgttaat aattcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
    10261 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
    10321 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
    10381 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
    10441 cgccatgcat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
    10501 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
    10561 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
    10621 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
    10681 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
    10741 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
    10801 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
    10861 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
    10921 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
    10981 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
    11041 tcagagcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
    11101 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct
    11161 gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt
    11221 ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt
    11281 cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa gatccttttt
    11341 gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg
    11401 tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg
    11461 gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt
    11521 tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc
    11581 cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt
    11641 ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg ggaagggcga
    11701 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
    11761 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
    11821 ttgtaatacg actcactata gggcgaattg aatttagcgg ccgcgaattc gccctt
//
