LOCUS       sequence-405449-       12581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac-based transposon vector plasmid which encodes the 
            expression units of liver-enriched transcription factor genes 
            (hHNF1-alpha-hHNF3-beta-hCEBP-alpha-hCEBP-beta-hCEBP-gamma) under 
            control of the TRE/PCMVmin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12581)
  AUTHORS   Kitano H, Kawabe Y, Kamihira M
  TITLE     HepG2-Based Designer Cells with Heat-Inducible Enhanced Liver 
            Functions.
  JOURNAL   Cells. 2022 Apr 1;11(7):1194. doi: 10.3390/cells11071194.
  PUBMED    35406758
REFERENCE   2  (bases 1 to 12581)
  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: "Cells."; 
            date: "2022-04-1"; volume: "11(7):1194. doi"; pages: " 
            10.3390/cells11071194"
COMMENT     Sequence Label: 
            PB513B-1/TRE-hHNF1-alpha-hHNF3-beta-hCEBP-alpha-hCEBP-beta-hCEBP-gam
            ma-EF1-alpha-Puro
FEATURES             Location/Qualifiers
     source          1..12581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1368..1386
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1404..1422
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1439..1457
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1475..1493
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1511..1529
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1546..1564
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1582..1600
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1612..1650
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1647..1671
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1689..1698
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3597..3653
                     /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             5058..5111
                     /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             6195..6251
                     /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             7296..7352
                     /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"
     promoter        7844..8055
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             8068..8336
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      8367..8376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8379..8390
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             8382..8435
                     /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             8436..9035
                     /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(8436..8455)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8932..8952
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(9089..9109)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    9151..9272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9188..9207)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9242..9261
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   10035..10069
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10560..10576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10560..10576)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10573..10595)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10584..10600
                     /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(10608..10614,10615..10632,10633..10638))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10653..10674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10791..10808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10962..11550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11042..11061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11721..12512,12513..12581))
                     /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     12344..12363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccacta gttttactcc ctatcagtga
     1381 tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgatgtc gagtttactc
     1441 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg
     1501 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttatccct atcagtgata
     1561 gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg aggtaggcgt
     1621 gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat cgcctggaat
     1681 ctagagccgc caccatggtg tccaagctgt cccagctgca aaccgagctg ctggccgctc
     1741 tgctggagag cggactgagc aaagaggccc tgatccaggc cctcggcgaa cccggacctt
     1801 atctgctggc cggcgaaggc cctctggaca aaggcgagag ctgtggcgga ggcagaggcg
     1861 aactggctga gctgcccaat ggactgggag agaccagggg cagcgaggac gagacagacg
     1921 acgacggcga ggactttacc cctcccatcc tgaaagagct ggagaacctg agccccgaag
     1981 aggccgccca tcagaaagcc gtggtggaga cactgctgca ggaggacccc tggagggtcg
     2041 ccaagatggt gaagagctac ctgcagcagc acaacatccc ccaaagagaa gtggtggaca
     2101 ccaccggcct gaatcaaagc cacctgtccc agcacctgaa taagggcacc cccatgaaaa
     2161 cccagaagag ggccgctctc tacacctggt acgtgagaaa gcagagggaa gtggctcagc
     2221 agtttaccca tgccggccaa ggcggcctga tcgaagagcc caccggcgat gagctgccta
     2281 ccaagaaggg aaggaggaac aggttcaagt ggggacctgc ctcccagcag atcctgtttc
     2341 aggcctacga gaggcagaag aaccctagca aagaggagag ggagaccctg gtggaagagt
     2401 gcaacagagc cgaatgcatc cagaggggcg tcagccctag ccaggctcag ggactgggca
     2461 gcaatctggt caccgaagtg agggtgtaca actggttcgc caacaggaga aaggaggagg
     2521 ccttcagaca caagctggct atggatacct acagcggacc tcctcctgga cctggacctg
     2581 gacccgctct gcctgcccat tccagccccg gactgcctcc tcccgccctg agcccctcca
     2641 aagtgcatgg agtgaggtac ggccaacctg ccaccagcga gaccgctgag gtgcctagct
     2701 ccagcggagg acctctggtg accgtctcca cccccctgca tcaggtgagc cctacaggac
     2761 tggagcccag ccacagcctg ctgagcaccg aggctaagct ggtgtccgct gccggaggac
     2821 ctctccctcc tgtgtccacc ctgaccgccc tccacagcct ggaacagacc tcccccggcc
     2881 tgaaccaaca acctcagaac ctgatcatgg ccagcctgcc cggagtgatg accatcggac
     2941 ctggcgagcc tgcttccctg ggccccacat ttaccaacac aggagcctcc acactggtga
     3001 tcggcctggc ttccacccag gcccagtccg tgcctgtgat caacagcatg ggctccagcc
     3061 tgaccacact ccaacccgtg caattcagcc agcctctgca cccctcctac cagcagcccc
     3121 tgatgccccc tgtgcagtcc cacgtcaccc aatccccctt catggccaca atggctcagc
     3181 tgcagtcccc ccatgccctg tattcccaca agcccgaggt ggctcaatac acccacacag
     3241 gactgctgcc tcagaccatg ctgatcaccg acaccaccaa cctgtccgcc ctggccagcc
     3301 tgacacccac aaagcaggtc tttaccagcg acaccgaggc cagctccgaa agcggcctcc
     3361 acacccctgc ttcccaggct accaccctgc acgtgcccag ccaagatcct gccggaatcc
     3421 agcacctgca acctgctcac aggctgagcg cttcccctac cgtgagctcc tccagcctgg
     3481 tcctgtacca gtccagcgac agctccaatg gccaatccca cctgctgccc agcaaccata
     3541 gcgtcatcga gaccttcatc agcacccaga tggcctcctc cagccaggga tctggcgcca
     3601 ccaacttcag cctgctgaaa caggctggcg acgtggaaga gaaccccgga cctatgcaca
     3661 gcgcctcttc tatgctgggc gccgtgaaga tggaaggcca cgagccatct gactggtcca
     3721 gctactatgc cgagcctgag ggctacagca gcgtgtccaa tatgaatgcc ggcctgggca
     3781 tgaacggcat gaatacctac atgagcatga gcgccgctgc catgggctct ggctctggaa
     3841 atatgtccgc cggcagcatg aacatgagca gctatgttgg cgccggaatg agcccatctc
     3901 tggccggaat gtctcctggt gctggtgcca tggctggcat gggaggatct gctggtgctg
     3961 ctggcgttgc cggaatggga cctcatctga gtcctagcct gtctccactc ggaggacaag
     4021 ctgcaggcgc tatgggagga ctggcccctt acgccaacat gaatagcatg agccctatgt
     4081 acggccaggc cggacttagc agagctagag atcccaagac ctaccggcgg agctacacac
     4141 acgccaagcc tccatacagc tacatcagcc tgatcaccat ggccatccag cagagcccca
     4201 acaagatgct gaccctgagc gaaatctacc agtggatcat ggatctgttc ccattctacc
     4261 ggcagaacca gcagcggtgg cagaacagca ttagacacag cctgagcttc aacgactgct
     4321 tcctgaaggt gccaagaagc cccgacaagc ctggcaaggg cagcttttgg acactgcacc
     4381 ccgacagcgg caacatgttc gagaatggct gctacctgcg gcggcagaaa cggttcaagt
     4441 gcgagaaaca gctggccctg aaagaggcag ctggcgctgc aggatctgga aagaaagctg
     4501 ctgctggggc ccaagcttct caggctcaac ttggagaagc tgccggacct gcctctgaaa
     4561 caccagccgg aacagagagc cctcacagct ctgcctctcc atgccaagag cacaagagag
     4621 gcggactggg agagctgaag ggaactcctg ctgctgcact gtctcctcca gagcctgctc
     4681 catctcctgg acagcagcaa caggctgctg ctcatctgct gggacctcca catcatcctg
     4741 gactgcctcc agaggctcac ctgaagcctg aacaccacta cgccttcaat caccccttca
     4801 gcatcaacaa cctgatgtcc agcgaacagc agcatcacca cagccaccac caccatcagc
     4861 ctcacaagat ggacctgaag gcctatgagc aagtgatgca ctaccccggc tacggctctc
     4921 ctatgcctgg atctcttgcc atgggacccg tgaccaacaa gaccggcctt gatgcttctc
     4981 ctctggccgc cgacacaagc tactatcagg gcgtgtacag cagacccatc atgaacagct
     5041 ccgaattcgg cagcggcgaa ggcagaggca gccttcttac atgtggcgac gtggaagaga
     5101 accccggacc tatggaaagc gccgacttct atgaggccga gccaagacct cctatgagca
     5161 gccatctgca gagccctcca cacgctccta gctctgccgc ctttggattt cctagaggcg
     5221 ctggacctgc tcagcctcct gctcctccag ctgctcctga acctctcggc ggaatttgcg
     5281 agcacgaaac cagcatcgac atcagcgcct acatcgaccc agccgccttc aacgatgagt
     5341 tcctggccga tctgttccag cacagccggc agcaagagaa ggccaaagct gctgttggac
     5401 ctactggtgg tggcggcgga ggcgattttg attatcctgg tgctcctgcc ggacctggcg
     5461 gagctgttat gccaggcgga gcacatggac ctccacctgg atatggatgt gccgccgctg
     5521 gctatctgga cggcagactg gaacctctgt acgagagagt tggagcccct gctctgaggc
     5581 ccctggtcat taagcaagag cccagagaag aggacgaggc caaacagctt gctctggccg
     5641 gactgttccc ttaccaacct cctccacctc caccaccttc tcaccctcat cctcatccgc
     5701 ctccagctca tctggctgct cctcatctgc agttccagat cgcccactgt ggccagacca
     5761 ccatgcatct gcaacccggc catcctacac caccacctac acctgtgcct tctccacatc
     5821 ctgctccagc tcttggagct gctggacttc ctggacctgg ctctgctctg aaaggactgg
     5881 gagccgctca tcccgatctg agagcatctg gtggatctgg cgccggaaag gccaagaaaa
     5941 gcgtggacaa gaacagcaac gagtacagag tgcggagaga gcggaacaat atcgccgtgc
     6001 gcaagagcag agacaaggcc aagcagcgga acgtggaaac ccagcagaag gtgctcgagc
     6061 tgaccagcga caacgaccgg ctgagaaaga gagtggaaca gctgagcaga gagctggaca
     6121 ccctgcgggg catctttaga cagctgcctg agagcagcct ggtcaaggcc atgggaaatt
     6181 gtgccggaag cggcgccacc aacttcagcc tgcttaaaca ggccggcgac gtcgaagaaa
     6241 atcctgggcc tatgcagcgg ctggtggctt gggaccctgc ttgtttgcct ttgccaccgc
     6301 cacctccagc cttcaagtct atggaagtgg ccaacttcta ctacgaggcc gactgtctgg
     6361 ccgctgccta tggtggaaaa gcagctcctg ctgcaccacc agctgctaga cctggaccta
     6421 gacctcctgc tggcgagctg ggatctatcg gcgatcacga gagagccatc gacttcagcc
     6481 cttacctgga accactgggc gctcctcaag ctcctgcacc agctacagcc accgatacct
     6541 ttgaagccgc tccaccagca ccggctccgg ctccagcttc ttctggacag caccacgact
     6601 ttctgagcga cctgttcagc gacgactacg gcggcaagaa ctgcaagaag cctgccgagt
     6661 acggctacgt gtcactgggc agattgggag ctgctaaggg cgctctgcac cctggatgtt
     6721 ttgcccctct tcaccctcca ccaccaccac cgcctccacc agctgaactg aaagccgagc
     6781 ctggattcga gcccgccgac tgcaagagaa aagaagaagc aggcgctcct ggcggaggtg
     6841 ctggaatggc tgcaggcttt ccttacgctc tgagagccta cctgggctac caggctgtgc
     6901 ctagcggatc ttctggcagc ctgagcacaa gcagcagctc tagccctcct ggcacacctt
     6961 ctccagccga tgctaaagcc cctcctacag cctgttatgc tggcgcagct cccgctccaa
     7021 gccaagtgaa gtccaaggcc aaaaagaccg tggataagca ctccgacgag tacaagatcc
     7081 ggcgcgagag aaacaacatt gccgtgcgga agtctcgcga caaagccaag atgagaaacc
     7141 tggaaacaca gcacaaagtc ctggaactga ccgccgagaa cgagagactg cagaagaagg
     7201 ttgagcagct gagccgggaa ctgagcaccc tgaggaacct gttcaagcag ctgccagagc
     7261 ctctgctggc ctcttctggc cattgtggat caggggccac aaatttcagt ctgctgaagc
     7321 aggcagggga tgttgaagag aatccaggac caatgagcaa gatcagccag cagaacagca
     7381 cccctggcgt gaacggcatc agcgtgatcc atacacaggc ccacgcctct ggactgcagc
     7441 aagttcctca actggtgcct gctggaccag gtggcggagg aaaagctgtt gcccctagca
     7501 agcagagcaa gaaatctagc cccatggacc ggaacagcga cgaataccgt cagcggagag
     7561 aaagaaacaa tatggctgtg aagaagtccc gcctcaagtc caagcagaaa gcccaggata
     7621 ccctgcagcg cgtgaaccag ctcaaagagg aaaacgagcg gctggaagcc aaaatcaagc
     7681 tgctgaccaa agaactgagc gtgctgaagg acctgttcct ggaacacgcc cacaacctgg
     7741 ccgacaacgt gcagagcatc agcaccgaga ataccaccgc cgacggcgat aatgccggcc
     7801 aatgagctag caaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     7861 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     7921 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     7981 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     8041 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     8101 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     8161 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     8221 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     8281 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag
     8341 ctgtgaccgg cgcctacgga tccgccgcca ccatggggat cgagggcaga ggaagtcttc
     8401 taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag cccacggtgc
     8461 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     8521 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     8581 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     8641 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     8701 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     8761 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     8821 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     8881 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     8941 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     9001 gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat tacaaaattt
     9061 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     9121 ctttaatgcc tttgtatcat gcgttaacta aacttgttta ttgcagctta taatggttac
     9181 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     9241 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggaattgact caaatgatgt
     9301 caattagtct atcagaagct catctggtct cccttccggg ggacaagaca tccctgttta
     9361 atatttaaac agcagtgttc ccaaactggg ttcttatatc ccttgctctg gtcaaccagg
     9421 ttgcagggtt tcctgtcctc acaggaacga agtccctaaa gaaacagtgg cagccaggtt
     9481 tagccccgga attgactgga ttcctttttt agggcccatt ggtatggctt tttccccgta
     9541 tccccccagg tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc
     9601 gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg
     9661 agcgccggac cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac
     9721 gtaattacat ccctgggggc tttggggggg ggctgtccct gatatctata acaagaaaat
     9781 atatatataa taagttatca cgtaagtaga acatgaaata acaatataat tatcgtatga
     9841 gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca cgcggtcgtt
     9901 atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg agctccaagc
     9961 ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa agagagagca
    10021 atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggt taatctagct
    10081 gcatcaggat catatcgtcg ggtctttttt ccggctcagt catcgcccaa gctggcgcta
    10141 tctgggcatc ggggaggaag aagcccgtgc cttttcccgc gaggttgaag cggcatggaa
    10201 agagtttgcc gaggatgact gctgctgcat tgacgttgag cgaaaacgca cgtttaccat
    10261 gatgattcgg gaaggtgtgg ccatgcacgc ctttaacggt gaactgttcg ttcaggccac
    10321 ctgggatacc agttcgtcgc ggcttttccg gacacagttc cggatggtca gcccgaagcg
    10381 catcagcaac ccgaacaata ccggcgacag ccggaactgc cgtgccggtg tgcagattaa
    10441 tgacagcggt gcggcgctgg gatattacgt cagcgaggac gggtatcctg gctggatgcc
    10501 gcagaaatgg acatggatac cccgtgagtt acccggcggg cgcgcttggc gtaatcatgg
    10561 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
    10621 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
    10681 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
    10741 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
    10801 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
    10861 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
    10921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
    10981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
    11041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
    11101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
    11161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
    11221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
    11281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
    11341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
    11401 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
    11461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
    11521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
    11581 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
    11641 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
    11701 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
    11761 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
    11821 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
    11881 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    11941 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    12001 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    12061 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
    12121 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    12181 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    12241 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    12301 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    12361 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    12421 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    12481 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    12541 aaaaagggaa taagggcgac acggaaatgt tgaatactca t
//
