LOCUS       sequence-271450-        8722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac vector for expression of 1x gRNA targeting POGZ 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: pPN461
FEATURES             Location/Qualifiers
     source          1..8722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(106..174,175..966)
                     /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(324..343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1137..1725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1626..1645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1879..1896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2013..2034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2049..2054,2055..2072,2073..2079)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2087..2103
                     /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     2111..2127
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2148..2166
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2315..2349
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        2728..3107
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3109..3386
                     /label=chicken beta-actin promoter
     intron          3387..4403
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4327..4349
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4411..4430
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(4481..4514)
                     /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      4520..4529
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5204..5257
                     /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             5258..5656
                     /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(5726..5746)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    5780..5901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5817..5836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5871..5890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5923..5956)
                     /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    5989..6013
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        6053..6301
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6053..6073
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6224..6243
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6323..6398
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(6484..6789)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     6654..6670
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6680..6698
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6746..6764)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6785..6801)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6809..6825
                     /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(6833..6839,6840..6857,6858..6863))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6878..6899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(6986..7010)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7106..7125)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7171..7226
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7225..7244)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   7930..7992
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(8059..8077)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8084..8100)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      8242..8697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8329..8348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8539..8560
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
       61 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1261 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1741 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1861 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     1921 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     1981 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     2041 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     2101 caatttcaca caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac
     2161 taaagggaac aaaagctggt acctcgcgcg acttggtttg ccattcttta gcgcgcgtcg
     2221 cgtcacacag cttggccaca atgtggtttt tgtcaaacga agattctatg acgtgtttaa
     2281 agtttaggtc gagtaaagcg caaatctttt ttaaccctag aaagatagtc tgcgtaaaat
     2341 tgacgcatgc attcttgaaa tattgctctc tctttctaaa tagcgcgaat ccgtcgctgt
     2401 gcatttagga catctcagtc gccgcttgga gctcccgtga ggcgtgcttg tcaatgcggt
     2461 aagtgtcact gattttgaac tataacgacc gcgtgagtca aaatgacgca tgattatctt
     2521 ttacgtgact tttaagattt aactcatacg ataattatat tgttatttca tgttctactt
     2581 acgtgataac ttattatata tatattttct tgttatagat atcgtgacta atatataata
     2641 aaatgggtag ttctttagac gatgagcata tcctctctgc tcttctgcaa agcgatgacg
     2701 agcttgttgg ctagcggtgg cggcctcgac attgattatt gactagttat taatagtaat
     2761 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     2821 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     2881 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     2941 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     3001 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     3061 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
     3121 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
     3181 ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag
     3241 gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca
     3301 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
     3361 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc
     3421 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
     3481 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc
     3541 ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg
     3601 gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct
     3661 ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca
     3721 gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg
     3781 ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt
     3841 cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt
     3901 cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc
     3961 ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg
     4021 ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc
     4081 ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc
     4141 cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg
     4201 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     4261 tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca
     4321 gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc
     4381 atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc
     4441 attttggcaa agaattcgcg gccgcggtac cggcgcgccg gaagttccta tactttctag
     4501 agaataggaa cttcatatcg ccaccatggc cagctccgag gatgtcatca aagagtttat
     4561 gagatttaag gtcaagatgg agggaagcgt caacggacac gagttcgaga ttgagggaga
     4621 aggagaaggc cggccttacg agggcacaca aaccgctaag ctcaaggtca caaaaggagg
     4681 acccctcccc ttctcctggg atattctgag ccctcagttc cagtacggaa gcaaagccta
     4741 tgttaaacac cctgccgaca tccctgacta tctgaagctc tccttccctg aaggcttcaa
     4801 gtgggagaga ttcatgaact tcgaggacgg aggcgtggtg acagtcacac aagatagcac
     4861 cctccaggac ggagagttta tttataaggt gaaactcaga ggaaccaact tcccctccga
     4921 tggccctgtc atgcaaaaaa aaacaatggg atgggaagcc tccaccgaga gaatgtatcc
     4981 tgaggatggc gctctgaaag gcgaaattaa aatgagactg aaactcaaag acggaggaca
     5041 ctacgatgcc gaggtcaaaa caacctacaa ggccaagaaa caagtgcagc tgcctggcgc
     5101 ctacatgact gatattaaac tcgacattat cagccataat ggggactaca ccatcgtgga
     5161 acaatatgag agagctgagg gcagacatag cacaggcgct ggagagggca gaggaagtct
     5221 tctaacatgc ggtgacgtgg aggagaatcc cggccctatg gccaagcctt tgtctcaaga
     5281 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     5341 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     5401 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     5461 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     5521 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     5581 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     5641 tgtgtgggag ggctaaaatc aacctctgga cgaaattccg cggttattac aaaatttgtg
     5701 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     5761 taatgccttt gtatcagtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     5821 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5881 caaactcatc aatgtatctt atcatgtctg gaattgactc aagaagttcc tatactttct
     5941 agagaatagg aacttcatga tgtggcgcgc cggatcctgc aggttatcac aagtttgtac
     6001 aaaaaagcag gctttaaagg aaccgccttt tgctggcctt ttgctcacat gtgagggcct
     6061 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg
     6121 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
     6181 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
     6241 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
     6301 cgtcctcggg tcagactacc gggttttaga gctagaaata gcaagttaaa ataaggctag
     6361 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta gagctagaaa
     6421 tagcaagtta aaataaggct agtccgtttt tagcgcgtgc gccaattctg cagacaaatg
     6481 gctctagaga ccggcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     6541 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6601 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     6661 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta ccgggccccc
     6721 cctcgaggtc ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     6781 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6841 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6901 ttgcgttgcg ctcactgccc gctttccacc ggtggtctct ctagaggtac ccgttacata
     6961 acttacggta aatggcccga tctagaccca gctttcttgt acaaagtggt gataactcta
     7021 gagaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     7081 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     7141 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     7201 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     7261 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     7321 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     7381 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     7441 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     7501 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     7561 tgcagcccaa gcttggatcc ctcgagttaa ttaacgagag cataatattg atatgtgcca
     7621 aagttgtttc tgactgacta ataagtataa tttgtttcta ttatgtatag gttaagctaa
     7681 ttacttattt tataatacaa catgactgtt tttaaagtac aaaataagtt tatttttgta
     7741 aaagagagaa tgtttaaaag ttttgttact ttatagaaga aattttgagt ttttgttttt
     7801 ttttaataaa taaataaaca taaataaatt gtttgttgaa tttattatta gtatgtaagt
     7861 gtaaatataa taaaacttaa tatctattca aattaataaa taaacctcga tatacagacc
     7921 gataaaacac atgcgtcaat tttacgcatg attatcttta acgtacgtca caatatgatt
     7981 atctttctag ggttaaataa tagtttctaa tttttttatt attcagcctg ctgtcgtgaa
     8041 taccgagctc caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
     8101 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     8161 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     8221 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     8281 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     8341 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     8401 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     8461 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     8521 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     8581 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     8641 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     8701 tggcactttt cggggaaatg tg
//
