LOCUS       sequence-271449-        8722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac vector for expression of 1x gRNA targeting TCF4 for 
            CRISPRi; mRFP-T2A-BlasticidinR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8722)
  AUTHORS   Hazelbaker DZ, Beccard A, Angelini G, Mazzucato P, Messana A, Lam D,
            Eggan K, Barrett LE
  TITLE     A multiplexed gRNA piggyBac transposon system facilitates efficient 
            induction of CRISPRi and CRISPRa in human pluripotent stem cells.
  JOURNAL   Sci Rep. 2020 Jan 20;10(1):635. doi: 10.1038/s41598-020-57500-1.
  PUBMED    31959800
REFERENCE   2  (bases 1 to 8722)
  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: "Sci Rep."; 
            date: "2020-01-20"; volume: "10(1):635. doi"; pages: " 
            10.1038/s41598-020-57500-1"
COMMENT     Sequence Label: pPN459
FEATURES             Location/Qualifiers
     source          1..8722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(131..199,200..991)
                     /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(349..368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1162..1750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1651..1670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1904..1921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2074..2079,2080..2097,2098..2104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2128
                     /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     2136..2152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2173..2191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2340..2374
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        2753..3132
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3134..3411
                     /label=chicken beta-actin promoter
     intron          3412..4428
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4352..4374
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4436..4455
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(4506..4539)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      4545..4554
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5229..5282
                     /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             5283..5681
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(5751..5771)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    5805..5926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5842..5861)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5896..5915
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5948..5981)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    6014..6038
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        6078..6326
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6078..6098
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6249..6268
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6348..6423
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(6509..6814)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     6679..6695
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6705..6723
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6771..6789)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6810..6826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6834..6850
                     /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(6858..6864,6865..6882,6883..6888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6903..6924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(7011..7035)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7131..7150)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7196..7251
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7250..7269)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   7955..8017
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(8084..8102)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8109..8125)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      8267..8695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8354..8373)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8564..8585
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2161 cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggtacctc gcgcgacttg
     2221 gtttgccatt ctttagcgcg cgtcgcgtca cacagcttgg ccacaatgtg gtttttgtca
     2281 aacgaagatt ctatgacgtg tttaaagttt aggtcgagta aagcgcaaat cttttttaac
     2341 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
     2401 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
     2461 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg
     2521 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat
     2581 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta
     2641 tagatatcgt gactaatata taataaaatg ggtagttctt tagacgatga gcatatcctc
     2701 tctgctcttc tgcaaagcga tgacgagctt gttggctagc ggtggcggcc tcgacattga
     2761 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     2821 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     2881 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     2941 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     3001 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     3061 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     3121 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     3181 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     3241 gggggggggg gggggcgcgc gccrggsggg gcggggcggg gcgaggggcg gggcggggcg
     3301 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
     3361 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
     3421 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
     3481 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     3541 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
     3601 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
     3661 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     3721 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
     3781 ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg
     3841 gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc
     3901 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
     3961 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     4021 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
     4081 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
     4141 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
     4201 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     4261 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     4321 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     4381 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     4441 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgcggccgc ggtaccggcg
     4501 cgccggaagt tcctatactt tctagagaat aggaacttca tatcgccacc atggccagct
     4561 ccgaggatgt catcaaagag tttatgagat ttaaggtcaa gatggaggga agcgtcaacg
     4621 gacacgagtt cgagattgag ggagaaggag aaggccggcc ttacgagggc acacaaaccg
     4681 ctaagctcaa ggtcacaaaa ggaggacccc tccccttctc ctgggatatt ctgagccctc
     4741 agttccagta cggaagcaaa gcctatgtta aacaccctgc cgacatccct gactatctga
     4801 agctctcctt ccctgaaggc ttcaagtggg agagattcat gaacttcgag gacggaggcg
     4861 tggtgacagt cacacaagat agcaccctcc aggacggaga gtttatttat aaggtgaaac
     4921 tcagaggaac caacttcccc tccgatggcc ctgtcatgca aaaaaaaaca atgggatggg
     4981 aagcctccac cgagagaatg tatcctgagg atggcgctct gaaaggcgaa attaaaatga
     5041 gactgaaact caaagacgga ggacactacg atgccgaggt caaaacaacc tacaaggcca
     5101 agaaacaagt gcagctgcct ggcgcctaca tgactgatat taaactcgac attatcagcc
     5161 ataatgggga ctacaccatc gtggaacaat atgagagagc tgagggcaga catagcacag
     5221 gcgctggaga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc
     5281 ctatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     5341 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     5401 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     5461 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     5521 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     5581 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     5641 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta aaatcaacct ctggacgaaa
     5701 ttccgcggtt attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     5761 tttacgctat gtggatacgc tgctttaatg cctttgtatc agttaacttg tttattgcag
     5821 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5881 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggaatt
     5941 gactcaagaa gttcctatac tttctagaga ataggaactt catgatgtgg cgcgccggat
     6001 cctgcaggtt atcacaagtt tgtacaaaaa agcaggcttt aaaggaaccg ccttttgctg
     6061 gccttttgct cacatgtgag ggcctatttc ccatgattcc ttcatatttg catatacgat
     6121 acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag atattagtac
     6181 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
     6241 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
     6301 atatcttgtg gaaaggacga aacaccgtaa acttgttcca agtttaggtt ttagagctag
     6361 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     6421 tgcttttttg ttttagagct agaaatagca agttaaaata aggctagtcc gtttttagcg
     6481 cgtgcgccaa ttctgcagac aaatggctct agagaccggc gccgctacag ggcgcgtccc
     6541 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6601 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6661 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     6721 agggcgaatt gggtaccggg ccccccctcg aggtcctcca gcttttgttc cctttagtga
     6781 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6841 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6901 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccaccggtgg
     6961 tctctctaga ggtacccgtt acataactta cggtaaatgg cccgatctag acccagcttt
     7021 cttgtacaaa gtggtgataa ctctagagaa ttcactcctc aggtgcaggc tgcctatcag
     7081 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     7141 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     7201 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     7261 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     7321 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     7381 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     7441 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     7501 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     7561 cttctcttat ggagatccct cgacctgcag cccaagcttg gatccctcga gttaattaac
     7621 gagagcataa tattgatatg tgccaaagtt gtttctgact gactaataag tataatttgt
     7681 ttctattatg tataggttaa gctaattact tattttataa tacaacatga ctgtttttaa
     7741 agtacaaaat aagtttattt ttgtaaaaga gagaatgttt aaaagttttg ttactttata
     7801 gaagaaattt tgagtttttg ttttttttta ataaataaat aaacataaat aaattgtttg
     7861 ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct attcaaatta
     7921 ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac gcatgattat
     7981 ctttaacgta cgtcacaata tgattatctt tctagggtta aataatagtt tctaattttt
     8041 ttattattca gcctgctgtc gtgaataccg agctccaatt cgccctatag tgagtcgtat
     8101 tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     8161 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     8221 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     8281 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     8341 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     8401 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     8461 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     8521 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     8581 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     8641 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     8701 aaaatattaa cgcttacaat tt
//
