LOCUS       sequence-386245-       10587 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac plasmid for stable expression of TIR1, mCherry, and 
            Puromycin resistance in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10587)
  AUTHORS   Spencley AL, Bar S, Swigut T, Flynn RA, Lee CH, Chen LF, Bassik MC, 
            Wysocka J
  TITLE     Co-transcriptional genome surveillance by HUSH is coupled to 
            termination machinery.
  JOURNAL   Mol Cell. 2023 May 18;83(10):1623-1639.e8. doi: 
            10.1016/j.molcel.2023.04.014. Epub 2023 May 9.
  PUBMED    37164018
REFERENCE   2  (bases 1 to 10587)
  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.molcel.2023.04.014"; journalName: "Mol Cell"; date: 
            "2023-05-18- 18"; volume: "83"; issue: "10"; pages: "1623-1639.e8"
COMMENT     Sequence Label: pAS_pb_Ef1a-TIR1_Ubc-mCherry-Puro
FEATURES             Location/Qualifiers
     source          1..10587
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(95..114)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      908..1496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1397..1416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1650..1667
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1784..1805
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1820..1825,1826..1843,1844..1850)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1858..1874
                     /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     1863..1885
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1882..1898
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1882..1898
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   2389..2423
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    3186..3307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3197..3216)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3251..3270
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3433..4032)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3513..3533)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     regulatory      4029..4038
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4045..4098)
                     /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(4108..4815)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     4652..4670
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(4826..6037)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(4970..4989)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     misc_feature    6044..6632
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6150..6161
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(6650..6691)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(6692..8416)
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     LTR             8441..8709
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(8722..8933)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    8980..9097
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     repeat_region   9528..9590
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9720..9737)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9720..9736)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9729..9751)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9878..10333
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9965..9984)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10175..10196
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10359..10463
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10464..10532,10533..10587,1..737)
                     /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"
ORIGIN
        1 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
       61 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
      121 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      181 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      241 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      301 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      361 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      421 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      481 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      541 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      601 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      661 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
      721 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
      781 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
      841 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      901 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      961 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1021 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     1081 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1141 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1201 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1261 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1321 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1381 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1441 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1501 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     1561 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     1621 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     1681 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     1741 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     1801 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     1861 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgccc
     1921 gccgggtaac tcacggggta tccatgtcca tttctgcggc atccagccag gatacccgtc
     1981 ctcgctgacg taatatccca gcgccgcacc gctgtcatta atctgcacac cggcacggca
     2041 gttccggctg tcgccggtat tgttcgggtt gctgatgcgc ttcgggctga ccatccggaa
     2101 ctgtgtccgg aaaagccgcg acgaactggt atcccaggtg gcctgaacga acagttcacc
     2161 gttaaaggcg tgcatggcca caccttcccg aatcatcatg gtaaacgtgc gttttcgctc
     2221 aacgtcaatg cagcagcagt catcctcggc aaactctttc catgccgctt caacctcgcg
     2281 ggaaaaggca cgggcttctt cctccccgat gcccagatag cgccagcttg ggcgatgact
     2341 gagccggaaa aaagacccga cgatatgatc ctgatgcagc tagattaacc ctagaaagat
     2401 agtctgcgta aaattgacgc atgcattctt gaaatattgc tctctctttc taaatagcgc
     2461 gaatccgtcg ctgtgcattt aggacatctc agtcgccgct tggagctccc gtgaggcgtg
     2521 cttgtcaatg cggtaagtgt cactgatttt gaactataac gaccgcgtga gtcaaaatga
     2581 cgcatgatta tcttttacgt gacttttaag atttaactca tacgataatt atattgttat
     2641 ttcatgttct acttacgtga taacttatta tatatatatt ttcttgttat agatatcagg
     2701 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
     2761 cagcgagccg cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca
     2821 gcgtgcgggg acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct
     2881 tctcgctgct ctttgagcct gcagacacct ggggggatac ggggaaaaag ccataccaat
     2941 gggccctaaa aaaggaatcc agtcaattcc ggggctaaac ctggctgcca ctgtttcttt
     3001 agggacttcg ttcctgtgag gacaggaaac cctgcaacct ggttgaccag agcaagggat
     3061 ataagaaccc agtttgggaa cactgctgtt taaatattaa acagggatgt cttgtccccc
     3121 ggaagggaga ccagatgagc ttctgataga ctaattgaca tcatttgagt caattccaga
     3181 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3241 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3301 acaagtttag ttagttaacg catgatacaa aggcattaaa gcagcgtatc cacatagcgt
     3361 aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt aatccagagg
     3421 ttgattgtcg acggcaccgg gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac
     3481 ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc
     3541 ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc cggggagcac
     3601 gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg tggccaggaa
     3661 ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc
     3721 cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc
     3781 ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg cgacccacac
     3841 cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc
     3901 gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg tagtcggcga acgcggcggc
     3961 gagggtgcgt acggcccggg ggacgtcgtc gcgggtggcg aggcgcaccg tgggcttgta
     4021 ctcggtggcc atggtggctc tagatgggcc aggattctcc tcgacgtcac cgcatgttag
     4081 cagacttcct ctgccctcac cggatccctt gtacagctcg tccatgccgc cggtggagtg
     4141 gcggccctcg gcgcgttcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat
     4201 gtccaacttg atgttgacgt tgtaggcgcc gggcagctgc acgggcttct tggccttgta
     4261 ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc agcttcagcc tctgcttgat
     4321 ctcgcccttc agggcgccgt cctcggggta catccgctcg gaggaggcct cccagcccat
     4381 ggttttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt
     4441 gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc
     4501 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct tcaagtagtc
     4561 ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact gaggggacag
     4621 gatgtcccag gcgaagggca gggggccacc cttggtcacc ttcagcttgg cggtctgggt
     4681 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga
     4741 gccctccatg tgcaccttga agcgcatgaa ctccttgatg atggccatgt tatcctcctc
     4801 gcccttgctc accatatccg ctagcgtcta acaaaaaagc caaaaacggc cagaatttag
     4861 cggacaattt actagtctaa cactgaaaat tacatattga cccaaatgat tacatttcaa
     4921 aaggtgccta aaaaacttca caaaacacac tcgccaaccc cgagcgcata gttcaaaacc
     4981 ggagcttcag ctacttaaga agataggtac ataaaaccga ccaaagaaac tgacgcctca
     5041 cttatccctc ccctcaccag aggtccggcg cctgtcgatt caggagagcc taccctaggc
     5101 ccgaaccctg cgtcctgcga cggagaaaag cctaccgcac acctaccggc aggtggcccc
     5161 accctgcatt ataagccaac agaacgggtg acgtcacgac acgacgaggg cgcgcgctcc
     5221 caaaggtacg ggtgcactgc ccaacggcac cgccataact gccgcccccg caacagacga
     5281 caaaccgagt tctccagtca gtgacaaact tcacgtcagg gtccccagat ggtgccccag
     5341 cccatctcac ccgaataaga gctttcccgc attagcgaag gcctcaagac cttgggttct
     5401 tgccgcccac catgcccccc accttgtttc aacgacctca cagcccgcct cacaagcgtc
     5461 ttccattcaa gactcgggaa cagccgccat tttgctgcgc tccccccaac ccccagttca
     5521 gggcaacctt gctcgcggac ccagactaca gcccttggcg gtctctccac acgcttccgt
     5581 cccaccgagc ggcccggcgg ccacgaaagc cccggccagc ccagcagccc gctactcacc
     5641 aagtgacgat cacagcgatc cacaaacaag aaccgcgacc caaatcccgg ctgcgacgga
     5701 actagctgtg ccacacccgg cgcgtcctta tataatcatc ggcgttcacc gccccacgga
     5761 gatccctccg cagaatcgcc gagaagggac tacttttcct cgcctgttcc gctctctgga
     5821 aagaaaacca gtgccctaga gtcacccaag tcccgtccta aaatgtcctt ctgctgatac
     5881 tggggttcta aggccgagtc ttatgagcag cgggccgctg tcctgagcgt ccgggcggaa
     5941 ggatcaggac gctcgctgcg cccttcgtct gacgtggcag cgctcgccgt gaggaggggg
     6001 gcgcccgcgg gaggcgccaa aacccggcgc ggaggccgga tccgcgggga ggcggcccaa
     6061 agggagatcc gactcgtctg agggcgaagg cgaagacgcg gaagaggccg cagagccggc
     6121 agcaggccgc gggaaggaag gtccgctgga ttgagggccg aagggacgta gcagaaggac
     6181 gtcccgcgca gaatccaggt ggcaacacag gcgagcagcc atggaaagga cgtcagcttc
     6241 cccgacaaca ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca
     6301 aggcaggcgg cgatgagttc cgccgtggca atagggaggg ggaaagcgaa agtcccggaa
     6361 aggagctgac aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg
     6421 cacaccacgc cacgttgcct gacaacgggc cacaactcct cataaagaga cagcaaccag
     6481 gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc
     6541 attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat
     6601 ctttcacaaa ttttgtaatc cagaggttga ttgtcgacgc ggccgcttag gtgctgtcca
     6661 ggcccagcag ggggttgggg atgggcttgc ccagaatttt cacgaaattg ggagcgtcat
     6721 ctcgtgcgcc agctgtggtc cgatagacgt acagtttctc cactttgggc agatcgccgt
     6781 ggttttcctc catttcatta ctaccgtcgc gttcgttgat gacctcaaca ttcagcattg
     6841 gcattttact agccaggact tggcatccct tgagggtcac gttgcagctt gacatccaaa
     6901 gtgacctcat cgtctcgtac cgggcgaaat tgcccagcag tgcggcgtcc ccaaaagggg
     6961 aatcgcgaat ttcgagtttt ctcaggttct tgcacccgtt catcacgtgc atcatccctt
     7021 tatctgaatc tccggcgaat gcgattgaca gcatttccag ctgcttggcg tacttcccaa
     7081 tatacatgaa cactttgtca gtcagcaggc cggatatgct cagccgctgg agacccttac
     7141 attcgcgtac tatggcaccg aacccttcgt ccagtggctg tgacgttacg acgtcgggct
     7201 taccgggctc caagatgcac agtctaaacc ttgtgaagtt tggacagttt ttggcaacgg
     7261 taacaagggc agcattggtc atctgatgac agaagtacag caaactattg agtttggggc
     7321 accccaggga cactgctacc aatccctcct cggtgactgc gctgtatccg gccacataga
     7381 aatctgaagg gaacaccctc agctcctgca gatctttgca ggaggaggcc acaacttgca
     7441 gtcccttgtc tgaaatacaa tccagtaccc agagcctttg gagcttcacg cagcggctga
     7501 tcatcttagt aaggtcacta gcgtcgaggg ttggagcgta actcagatta agtccggtca
     7561 actgtgcaca caggggatag ataaaagaca ggcagacagg gctagcgtcc caaaacccgc
     7621 tcagtgacct aagcattttg catttttcca gagcactagt gagcttaaag tagctctctg
     7681 tctggaaatc atcggtcaga tttccggtcc ccagatcctc caggttggga gtccggagca
     7741 aaatcttggc gagcgtgtcc acagaaacgg atctgttcag ccgcagagat ctcaggttgg
     7801 gggatcggga aacgagtctc tccaaactcc cggcattcac ttcgcccttg atacaagcga
     7861 aattcaagct caccaggcta gtgcagctat ccgggaaaca gctcagccac cgaggtcccc
     7921 tatcctctac ctcattttcc tgcaggtcca gttctcgcag cagtttacaa tggctagcta
     7981 cagcagccag accgtcggtg ctaaatcctt cacagctgat cagcaccagg gcgcggaatc
     8041 gggggaagct gcgggcaagc agttccagtg actcgtctga caccaccatc cttttcatcc
     8101 tgagttcttc cagaccatga cacccgcgcg cagcggcctc gatccagggc ccggcgtagc
     8161 cgccccagtc cggaggcacc aaattgaagt cggcaaagtg aggcttgccc ttcactgtaa
     8221 gggccctgac gttcgggaac cgggcggcca ctcgcccggc tcgtacggca taacaattac
     8281 ccacaaaaac acctctgcgg ctcaagcgtt cgatctcata ccagaccttg cacaccaggg
     8341 acacggtatt cctatctcgt tgggcgggaa ggaatgagaa aatatgttcg accacctcct
     8401 cagggaagta tgtcatgaat tcggcgccgg tcacagcttg gatctgtaac ggcgcagaac
     8461 agaaaacgaa acaaagacgt agagttgagc aagcagggtc aggcaaagcg tggagagccg
     8521 gctgagtcta ggtaggctcc aagggagcgc cggacaaagg cccggtctcg acctgagctt
     8581 taaacttacc tagacggcgg acgcagttca ggaggcacca caggcgggag gcggcagaac
     8641 gcgactcaac cggcgtggat ggcggcctca ggtagggcgg cgggcgcgtg aaggagagat
     8701 gcgagcccct cgaagcttca gctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt
     8761 cacggcgact actgcactta tatacggttc tcccccaccc tcgggaaaaa ggcggagcca
     8821 gtacacgaca tcactttccc agtttacccc gcgccacctt ctctaggcac ccgttcaatt
     8881 gccgacccct ccccccaact tctcggggac tgtgggcgat gtgcgctctg cccactgacg
     8941 ggcaccggat gatccctcga ggctagtctc gtgatcgata aaattttgaa tttttgtaat
     9001 ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc
     9061 tgcactgtac cccccaatcc ccccttttct tttaaaagtt ctagtctggc cgtggaggcc
     9121 ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg
     9181 aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg
     9241 gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc
     9301 gggggaggga cgtaattaca tccctggggg ctttgggggg gggctgtccc tcataaaagt
     9361 tttgttactt tatagaagaa attttgagtt tttgtttttt ttaataaata aataaacata
     9421 aataaattgt ttgttgaatt tattattagt atgtaagtgt aaatataata aaacttaata
     9481 tctattcaaa ttaataaata aacctcgata tacagaccga taaaacacat gcgtcaattt
     9541 tacacatgat tatctttaac gtacgtcaca atatgattat ctttctaggg ttaatctagc
     9601 tgcgtgttct gcagcgtgtc gagcatcttc atctgctcca tcacgctgta aaacacattt
     9661 gcaccgcgag tctgcccgtc ctccacgggt tcaaaaacgt gaatgaacga ggcgcgctca
     9721 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     9781 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     9841 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     9901 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     9961 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
    10021 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
    10081 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
    10141 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
    10201 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
    10261 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
    10321 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
    10381 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
    10441 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
    10501 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
    10561 atgctgaaga tcagttgggt gcacgag
//
