LOCUS       sequence-392528-       11545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of Expandable megakaryocyte cell lines from human iPS 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11545)
  AUTHORS   Nakamura S, Takayama N, Hirata S, Seo H, Endo H, Ochi K, Fujita K, 
            Koike T, Harimoto K, Dohda T, Watanabe A, Okita K, Takahashi N, 
            Sawaguchi A, Yamanaka S, Nakauchi H, Nishimura S, Eto K
  TITLE     Expandable megakaryocyte cell lines enable clinically applicable 
            generation of platelets from human induced pluripotent stem cells.
  JOURNAL   Cell Stem Cell. 2014 Apr 3;14(4):535-48. doi: 
            10.1016/j.stem.2014.01.011. Epub 2014 Feb 13.
  PUBMED    24529595
REFERENCE   2  (bases 1 to 11545)
  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"; doi: 
            "10.1016/j.stem.2014.01.011"; journalName: "Cell Stem Cell"; date: 
            "2014-04-3- 3"; volume: "14"; issue: "4"; pages: "535-48"
COMMENT     Sequence Label: CS-TRE-c-Myc-Ubc-rtTA
FEATURES             Location/Qualifiers
     source          1..11545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1059..1184
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1677..1910
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2095..2139
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2413..2530
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2572..2886
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2580..2598
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2616..2634
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2651..2669
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2687..2705
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2723..2741
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2758..2776
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2794..2812
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2859..2883
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2909..4228
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     promoter        4246..5457
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5294..5313
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5470..6087,6088..6093,6094..6129,6130..6132,6133..6168,
                     6169..6171,6172..6207,6208..6216)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    6240..6828
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6293..6313)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6711..6722)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7347..7527
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7553..7570)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7559..7783
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7829..8257
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7916..7935)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8126..8147
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8266..8286)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8271..8600
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8451..8586
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8513..8532
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8648..8695
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8714..9088
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    9218..9339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9255..9274)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9309..9328
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9388..9404)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9388..9404)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9401..9423)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9412..9428
                     /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(9436..9442,9443..9460,9461..9466))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9481..9502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9619..9636)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9790..10378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9870..9889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10549..11340,11341..11409))
                     /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     11172..11191
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11410..11514)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     1081 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     1141 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1201 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1261 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1321 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1381 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1441 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1501 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
     1561 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1621 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1681 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1741 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1801 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1861 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1921 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1981 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     2041 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     2101 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     2161 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     2221 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     2281 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     2341 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctac aaatggcagt
     2401 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     2461 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     2521 tcaaaatttt cgggtttatt acagggacag cagaaattca ctttgaattg cgagtttact
     2581 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
     2641 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
     2701 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
     2761 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
     2821 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
     2881 atcgcctgga gaattgcgta cggccagcat gcccctcaac gttagcttca ccaacaggaa
     2941 ctatgacctc gactacgact cggtgcagcc gtatttctac tgcgacgagg aggagaactt
     3001 ctaccagcag cagcagcaga gcgagctgca gcccccggcg cccagcgagg atatctggaa
     3061 gaaattcgag ctgctgccca ccccgcccct gtcccctagc cgccgctccg ggctctgctc
     3121 gccctcctac gttgcggtca cacccttctc ccttcgggga gacaacgacg gcggtggcgg
     3181 gagcttctcc acggccgacc agctggagat ggtgaccgag ctgctgggag gagacatggt
     3241 gaaccagagt ttcatctgcg acccggacga cgagaccttc atcaaaaaca tcatcatcca
     3301 ggactgtatg tggagcggct tctcggccgc cgccaagctc gtctcagaga agctggcctc
     3361 ctaccaggct gcgcgcaaag acagcggcag cccgaacccc gcccgcggcc acagcgtctg
     3421 ctccacctcc agcttgtacc tgcaggatct gagcgccgcc gcctcagagt gcatcgaccc
     3481 ctcggtggtc ttcccctacc ctctcaacga cagcagctcg cccaagtcct gcgcctcgca
     3541 agactccagc gccttctctc cgtcctcgga ttctctgctc tcctcgacgg agtcctcccc
     3601 gcagggcagc cccgagcccc tggtgctcca tgaggagaca ccgcccacca ccagcagcga
     3661 ctctgaggag gaacaagaag atgaggaaga aatcgatgtt gtttctgtgg aaaagaggca
     3721 ggctcctggc aaaaggtcag agtctggatc accttctgct ggaggccaca gcaaacctcc
     3781 tcacagccca ctggtcctca agaggtgcca cgtctccaca catcagcaca actacgcagc
     3841 gcctccctcc actcggaagg actatcctgc tgccaagagg gtcaagttgg acagtgtcag
     3901 agtcctgaga cagatcagca acaaccgaaa atgcaccagc cccaggtcct cggacaccga
     3961 ggagaatgtc aagaggcgaa cacacaacgt cttggagcgc cagaggagga acgagctaaa
     4021 acggagcttt tttgccctgc gtgaccagat cccggagttg gaaaacaatg aaaaggcccc
     4081 caaggtagtt atccttaaaa aagccacagc atacatcctg tccgtccaag cagaggagca
     4141 aaagctcatt tctgaagagg acttgttgcg gaaacgacga gaacagttga aacacaaact
     4201 tgaacagcta cggaactctt gtgcgtaaga attcgctagc gatctggcct ccgcgccggg
     4261 ttttggcgcc tcccgcgggc gcccccctcc tcacggcgag cgctgccacg tcagacgaag
     4321 ggcgcagcga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat
     4381 aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg
     4441 actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc
     4501 tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg
     4561 cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg
     4621 tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt
     4681 ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct
     4741 gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc
     4801 tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag
     4861 gtggggggca tggtgggcgg caagaaccca aggtcttgag gccttcgcta atgcgggaaa
     4921 gctcttattc gggtgagatg ggctggggca ccatctgggg accctgacgt gaagtttgtc
     4981 actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt
     5041 gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt
     5101 ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc
     5161 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     5221 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     5281 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     5341 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     5401 tgttagacta gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaag
     5461 cttgtcacca tgtctagact ggacaagagc aaagtcataa acggcgctct ggaattactc
     5521 aatggagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     5581 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc
     5641 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac
     5701 tttctgcgga acaacgccaa gtcattccgc tgtgctctcc tctcacatcg cgacggggct
     5761 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     5821 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     5881 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa
     5941 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc
     6001 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     6061 gagaaacagc taaagtgcga aagcggcggg ccggccgacg cccttgacga ttttgactta
     6121 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
     6181 cttgacgatt ttgaccttga catgctcccc gggtaactcg agggatcaag cttatcgata
     6241 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     6301 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     6361 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     6421 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     6481 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     6541 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     6601 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     6661 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6721 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6781 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     6841 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
     6901 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
     6961 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     7021 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct
     7081 accacacaca aggctacttc cctgattggc agaactacac accagggcca gggatcagat
     7141 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag
     7201 aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg
     7261 acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg
     7321 cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg
     7381 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     7441 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     7501 tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact
     7561 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     7621 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     7681 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     7741 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     7801 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     7861 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     7921 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     7981 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     8041 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     8101 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     8161 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     8221 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     8281 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     8341 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     8401 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     8461 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     8521 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     8581 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     8641 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     8701 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     8761 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     8821 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     8881 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     8941 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     9001 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     9061 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     9121 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     9181 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     9241 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     9301 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     9361 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     9421 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     9481 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     9541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     9601 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     9661 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     9721 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     9781 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     9841 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     9901 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     9961 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
    10021 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
    10081 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
    10141 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
    10201 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
    10261 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
    10321 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
    10381 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
    10441 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
    10501 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
    10561 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
    10621 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
    10681 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
    10741 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
    10801 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
    10861 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
    10921 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
    10981 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
    11041 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
    11101 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
    11161 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
    11221 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
    11281 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
    11341 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
    11401 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
    11461 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
    11521 tccccgaaaa gtgccacctg acgtc
//
