LOCUS       sequence-307963-        8909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggybacTransposon-based tunable and temporal expression control of 
            mutated beta-catenin and nuclear eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8909)
  AUTHORS   Chang Y, Syahirah R, Oprescu SN, Wang X, Jung J, Cooper SH, 
            Torregrosa-Allen S, Elzey BD, Hsu AY, Randolph LN, Sun Y, Kuang S, 
            Broxmeyer HE, Deng Q, Lian X, Bao X
  TITLE     Chemically-defined generation of human hemogenic endothelium and 
            definitive hematopoietic progenitor cells.
  JOURNAL   Biomaterials. 2022 May 6;285:121569. doi: 
            10.1016/j.biomaterials.2022.121569.
  PUBMED    35567999
REFERENCE   2  (bases 1 to 8909)
  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: 
            "Biomaterials."; date: "2022-05-6"; volume: "285:121569. doi"; 
            pages: " 10.1016/j.biomaterials.2022.121569"
COMMENT     Sequence Label: XLone-Puro beta-catenin P2A eGFP-NLS
FEATURES             Location/Qualifiers
     source          1..8909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..376
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    17..35
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    53..71
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    89..107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    125..143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    161..179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    197..215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    233..251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      383..392
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             434..469
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     CDS             2747..2803
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(2804..2806,2807..2809,2810..3520)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2804..2806,2807..2809,2810..3520)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2849..2870)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3110..3129)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3457..3478
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3521..3541
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3760..3894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   4170..4204
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4215..4232)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4215..4231)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4224..4246)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4440..4459)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4559..4581
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4619..4637)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4705..4809
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4810..4878,4879..5670)
                     /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(5028..5047)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5841..6429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6330..6349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6583..6600
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6717..6738
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6753..6758,6759..6776,6777..6783)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6791..6807
                     /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     6796..6818
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6815..6831
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6815..6831
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   6857..6919
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    7095..7176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7197..7796)
                     /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(7280..7300)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7777..7796
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(7806..7859)
                     /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             complement(7869..8612)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             complement(8613..8636)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      8636..8645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8665..8876)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 atcacctcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
       61 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
      121 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
      181 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
      241 agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa aagagtgctg
      301 attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
      361 ttcctaccct cgtaaaggta ccgccaccat ggctactcaa gctgacctga tggagttgga
      421 catggccatg gagccggaca gaaaagctgc tgtcagccac tggcagcagc agtcttactt
      481 ggatgctgga atccatgctg gtgccaccgc cacagctcct gccctgagtg gcaagggcaa
      541 ccctgaggaa gaagatgttg acacctccca agtcctttat gaatgggagc aaggcttttc
      601 ccagtccttc acgcaagagc aagtagctga tattgacggg cagtatgcaa tgactagggc
      661 tcagagggtc cgagctgcca tgttccctga gacgctagat gagggcatgc agatcccatc
      721 cacgcagttt gacgctgctc atcccactaa tgtccagcgc ttggctgaac catcacagat
      781 gttgaaacat gcagttgtca atttgattaa ctatcaggat gacgcggaac ttgccacacg
      841 tgcaattcct gagctgacaa aactgctaaa cgatgaggac caggtggtag ttaataaagc
      901 tgctgttatg gtccatcagc tttccaaaaa ggaagcttcc agacatgcca tcatgcgctc
      961 ccctcagatg gtgtctgcca ttgtacgcac catgcagaat acaaatgatg tagagacagc
     1021 tcgttgtact gctgggactc tgcacaacct ttctcaccac cgcgagggct tgctggccat
     1081 ctttaagtct ggtggcatcc cagcgctggt gaaaatgctt gggtcaccag tggattctgt
     1141 actgttctac gccatcacga cactgcataa tctcctgctc catcaggaag gagctaaaat
     1201 ggcagtgcgc ctagctggtg gactgcagaa aatggttgct ttgctcaaca aaacaaacgt
     1261 gaaattcttg gctattacaa cagactgcct tcagatctta gcttatggca atcaagagag
     1321 caagctcatc attctggcca gtggtggacc ccaagcctta gtaaacataa tgaggaccta
     1381 cacttatgag aagcttctgt ggaccacaag cagagtgctg aaagtgctgt ctgtctgctc
     1441 tagcaacaag ccggccattg tagaagctgg tgggatgcag gcactggggc ttcatctgac
     1501 agacccaagt cagcgacttg ttcaaaactg tctttggact ctcagaaacc tttcagatgc
     1561 agcgactaag caggaaggga tggaaggcct ccttgggact ctagtgcagc ttctgggttc
     1621 cgatgatata aatgtggtca cctgtgcagc tggaattctc tctaacctca cttgcaataa
     1681 ttacaaaaac aagatgatgg tgtgccaagt gggtggcata gaggctcttg tacgcaccgt
     1741 ccttcgtgct ggtgacaggg aagacatcac tgagcctgcc atctgtgctc ttcgtcatct
     1801 gaccagccgg catcaggaag ccgagatggc ccagaatgcc gttcgccttc attatggact
     1861 gcctgttgtg gttaaactcc tgcacccacc atcccactgg cctctgataa aggcaactgt
     1921 tggattgatt cgaaaccttg ccctttgccc agcaaatcat gcgcctttgc gggaacaggg
     1981 tgctattcca cgactagttc agctgcttgt acgagcacat caggacaccc aacggcgcac
     2041 ctccatgggt ggaacgcagc agcagtttgt ggagggcgtg cgcatggagg agatagtaga
     2101 agggtgtact ggagctctcc acatccttgc tcgggacgtt cacaaccgga ttgtaatccg
     2161 aggactcaat accattccat tgtttgtgca gttgctttat tctcccattg aaaatatcca
     2221 aagagtagct gcaggggtcc tctgtgaact tgctcaggac aaggaggctg cagaggccat
     2281 tgaagctgag ggagccacag ctcccctgac agagttactc cactccagga atgaaggcgt
     2341 ggcaacatac gcagctgctg tcctattccg aatgtctgag gacaagccac aggattacaa
     2401 gaagcggctt tcagtcgagc tgaccagttc cctcttcagg acagagccaa tggcttggaa
     2461 tgagactgca gatcttggac tggacattgg tgcccaggga gaagcccttg gatatcgcca
     2521 ggatgatccc agctaccgtt cttttcactc tggtggatac ggccaggatg ccttggggat
     2581 ggaccctatg atggagcatg agatgggtgg ccaccaccct ggtgctgact atccagttga
     2641 tgggctgcct gatctgggac acgcccagga cctcatggat gggctgcccc caggtgatag
     2701 caatcagctg gcctggtttg atactgacct ggctagcggc agcggcgcca ctaacttctc
     2761 cctgttgaaa caagcagggg atgtcgaaga gaatcccggg ccaatggtga gcaagggcga
     2821 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2881 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2941 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     3001 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3061 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     3121 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3181 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     3241 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     3301 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3361 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3421 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3481 cgccgccggg atcactctcg gcatggacga gctgtacaag ccaaagaaga agcgtaaggt
     3541 ataagtacaa gtaagaattc cgatcatatt caataaccca ctagtagacc acctcccctg
     3601 cgagctaagc tggacagcca atgacgggta agagagtgac atttttcact aacctaagac
     3661 aggagggccg tcagagctac tgcctaatcc aaagacgggt aaaagtgata aaaatgtatc
     3721 actccaacct aagacaggcg cagcttccga gggatttgag atccagacat gataagatac
     3781 attgatgagt ttggacaaac caaaactaga atgcagtgaa aaaaatgcct tatttgtgaa
     3841 atttgtgatg ctattgcctt atttgtaacc attataagct gcaataaaca agtttgatat
     3901 ctataacaag aaaatatata tataataagt tatcacgtaa gtagaacatg aaataacaat
     3961 ataattatcg tatgagttaa atcttaaaag tcacgtaaaa gataatcatg cgtcattttg
     4021 actcacgcgg tcgttatagt tcaaaatcag tgacacttac cgcattgaca agcacgcctc
     4081 acgggagctc caagcggcga ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt
     4141 tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga ctatctttct
     4201 agggttaaga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     4261 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     4321 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     4381 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     4441 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4501 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4561 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4621 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4681 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4741 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4801 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4861 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4921 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4981 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5041 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5101 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5161 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5221 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5281 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5341 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5401 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5461 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5521 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5581 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5641 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5701 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5761 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5821 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5881 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5941 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6001 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6061 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6121 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6181 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6241 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6301 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6361 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6421 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6481 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6541 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6601 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6661 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6721 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6781 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6841 gccaaggtcg acttaaccct agaaagataa tcatattgtg acgtacgtta aagataatca
     6901 tgcgtaaaat tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg
     6961 aatagatatt aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa
     7021 tttatttatg tttatttatt tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag
     7081 taacaaaact tttagcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     7141 ttatttgtaa ccattataag ctgcaataaa caagttaact aaacgggccc tctagatcag
     7201 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
     7261 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg
     7321 aaggcgggca ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc
     7381 agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc
     7441 ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg
     7501 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     7561 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     7621 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     7681 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     7741 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatctcg
     7801 agcctagggc cgggattctc ctccacgtca ccgcatgtta gaagacttcc tctgccctct
     7861 ccgctgcccc cggggagcat gtcaaggtca aaatcgtcaa gagcgtcagc aggcagcata
     7921 tcaaggtcaa agtcgtcaag ggcatcggct gggagcatgt ctaagtcaaa atcgtcaagg
     7981 gcgtcggtcg gcccgccgct ttcgcacttt agctgtttct ccaggccaca tatgattagt
     8041 tccaggccga aaaggaaggc aggttcggct ccctgccggt cgaacagctc aattgcttgt
     8101 ttcagaagtg ggggcataga atcggtggta ggtgtctctc tttcctcttt tgctacttga
     8161 tgctcctgtt cctccaatac gcagcccagt gtaaagtggc ccacggcgga cagagcgtac
     8221 agtgcgttct ccagggagaa gccttgctga cacaggaacg cgagctgatt ttccagggtt
     8281 tcgtactgtt tctctgttgg gcgggtgccg agatgcactt tagccccgtc gcgatgtgag
     8341 aggagagcac agcggtatga cttggcgttg ttccgcagaa agtcttgcca tgactcgcct
     8401 tccagggggc aggagtgggt atgatgcctg tccagcatct cgattggcag ggcatcgagc
     8461 agggcccgct tgttcttcac gtgccagtac agggtaggct gctcaactcc cagcttttga
     8521 gcgagtttcc ttgtcgtcag gccttcgata ccgactccat tgagtaattc cagagcagag
     8581 tttatgactt tgctcttgtc cagtctagac atcttatcgt catcgtcttt gtaatccatg
     8641 gtggcggatc ccgcgtcacg acacctgtgt tctggcggca aacccgttgc gaaaaagaac
     8701 gttcacggcg actactgcac ttatatacgg ttctccccca ccctcgggaa aaaggcggag
     8761 ccagtacacg acatcacttt cccagtttac cccgcgccac cttctctagg caccggttca
     8821 attgccgacc cctcccccca acttctcggg gactgtgggc gatgtgcgct ctgcccactg
     8881 acgggcaccg gagccactcg agtggaatt
//
