LOCUS       sequence-324885-        9402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac donor vector for expression of mRFP-T2A-BlasticidinR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9402)
  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 9402)
  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: pGEP163
FEATURES             Location/Qualifiers
     source          1..9402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..656
                     /label=chicken beta-actin promoter
     intron          657..1673
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1597..1619
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1681..1700
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1751..1784)
                     /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      1790..1799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2474..2527
                     /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             2528..2926
                     /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(2996..3016)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3050..3171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3087..3106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3141..3160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3193..3226)
                     /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    3259..3383
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3408..3413,3414..3431,3432..3438)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3492..4151
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3558..3577)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4493..4798
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4677..4696
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4839..4963)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5059..5078)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5124..5179
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5178..5197)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   5883..5945
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6011..6030)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6012..6030)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6037..6054)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6037..6053)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6046..6068)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6195..6650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6282..6301)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6492..6513
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6676..6780
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6781..6849,6850..7641)
                     /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(6999..7018)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7812..8400
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8301..8320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8554..8571
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8688..8709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8724..8729,8730..8747,8748..8754)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8762..8778
                     /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     8767..8789
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8786..8802
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8786..8802
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8823..8841
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   8990..9024
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggskrkggk cgcgcgccag gcggggcggg gcggggcgag gggcggggcg
      541 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      601 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      661 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      721 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      781 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      841 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg
      961 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg
     1021 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc
     1141 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg
     1201 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc
     1261 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
     1441 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
     1501 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     1561 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1621 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1681 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcgcg gccgcggtac
     1741 cggcgcgccg gaagttccta tactttctag agaataggaa cttcatatcg ccaccatggc
     1801 cagctccgag gatgtcatca aagagtttat gagatttaag gtcaagatgg agggaagcgt
     1861 caacggacac gagttcgaga ttgagggaga aggagaaggc cggccttacg agggcacaca
     1921 aaccgctaag ctcaaggtca caaaaggagg acccctcccc ttctcctggg atattctgag
     1981 ccctcagttc cagtacggaa gcaaagccta tgttaaacac cctgccgaca tccctgacta
     2041 tctgaagctc tccttccctg aaggcttcaa gtgggagaga ttcatgaact tcgaggacgg
     2101 aggcgtggtg acagtcacac aagatagcac cctccaggac ggagagttta tttataaggt
     2161 gaaactcaga ggaaccaact tcccctccga tggccctgtc atgcaaaaaa aaacaatggg
     2221 atgggaagcc tccaccgaga gaatgtatcc tgaggatggc gctctgaaag gcgaaattaa
     2281 aatgagactg aaactcaaag acggaggaca ctacgatgcc gaggtcaaaa caacctacaa
     2341 ggccaagaaa caagtgcagc tgcctggcgc ctacatgact gatattaaac tcgacattat
     2401 cagccataat ggggactaca ccatcgtgga acaatatgag agagctgagg gcagacatag
     2461 cacaggcgct ggagagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc
     2521 cggccctatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
     2581 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     2641 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     2701 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     2761 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     2821 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     2881 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaaaatc aacctctgga
     2941 cgaaattccg cggttattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3001 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcagtta acttgtttat
     3061 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3121 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3181 gaattgactc aagaagttcc tatactttct agagaatagg aacttcatga tgtggcgcgc
     3241 cggatcctgc aggttatcac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat
     3301 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     3361 acacaacata tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg
     3421 cttccggctc gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta
     3481 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     3541 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     3601 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     3661 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
     3721 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     3781 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     3841 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     3901 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     3961 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     4021 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     4081 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     4141 gcggggcgta aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc
     4201 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
     4261 gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
     4321 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
     4381 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
     4441 cggtttattg aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt
     4501 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     4561 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     4621 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     4681 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     4741 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     4801 gtcaggctcc cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg
     4861 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     4921 tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggtgataact ctagagaatt
     4981 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     5041 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     5101 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     5161 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     5221 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     5281 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     5341 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     5401 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     5461 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     5521 caagcttgga tccctcgagt taattaacga gagcataata ttgatatgtg ccaaagttgt
     5581 ttctgactga ctaataagta taatttgttt ctattatgta taggttaagc taattactta
     5641 ttttataata caacatgact gtttttaaag tacaaaataa gtttattttt gtaaaagaga
     5701 gaatgtttaa aagttttgtt actttataga agaaattttg agtttttgtt tttttttaat
     5761 aaataaataa acataaataa attgtttgtt gaatttatta ttagtatgta agtgtaaata
     5821 taataaaact taatatctat tcaaattaat aaataaacct cgatatacag accgataaaa
     5881 cacatgcgtc aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc
     5941 tagggttaaa taatagtttc taattttttt attattcagc ctgctgtcgt gaataccgag
     6001 ctccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     6061 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     6121 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     6181 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6241 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6301 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6361 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6421 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6481 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6541 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     6601 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     6661 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6721 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6781 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6841 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6901 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6961 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     7021 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     7081 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     7141 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     7201 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     7261 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7321 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7381 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7441 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7501 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7561 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7621 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7681 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7741 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7801 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7861 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7921 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7981 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     8041 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     8101 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     8161 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     8221 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8281 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8341 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8401 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8461 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     8521 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     8581 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     8641 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     8701 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     8761 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     8821 gaaattaacc ctcactaaag ggaacaaaag ctggtacctc gcgcgacttg gtttgccatt
     8881 ctttagcgcg cgtcgcgtca cacagcttgg ccacaatgtg gtttttgtca aacgaagatt
     8941 ctatgacgtg tttaaagttt aggtcgagta aagcgcaaat cttttttaac cctagaaaga
     9001 tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt ctaaatagcg
     9061 cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc cgtgaggcgt
     9121 gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg agtcaaaatg
     9181 acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat tatattgtta
     9241 tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta tagatatcgt
     9301 gactaatata taataaaatg ggtagttctt tagacgatga gcatatcctc tctgctcttc
     9361 tgcaaagcga tgacgagctt gttggctagc ggtggcggcc tc
//
