LOCUS       sequence-275218-        8722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac vector for expression of 1x gRNA targeting POGZ for 
            CRISPRa;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: pPN460
FEATURES             Location/Qualifiers
     source          1..8722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..659
                     /label=chicken beta-actin promoter
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1754..1787)
                     /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      1793..1802
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2477..2530
                     /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             2531..2929
                     /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(2999..3019)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3053..3174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3090..3109)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3144..3163
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3196..3229)
                     /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    3262..3286
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3326..3574
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3326..3346
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3497..3516
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3596..3671
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(3757..4062)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     3927..3943
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3953..3971
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4019..4037)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4058..4074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4082..4098
                     /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(4106..4112,4113..4130,4131..4136))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4151..4172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(4259..4283)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4379..4398)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4444..4499
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4498..4517)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   5203..5265
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(5332..5350)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5357..5373)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5515..5970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5602..5621)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5812..5833
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5996..6100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6101..6169,6170..6961)
                     /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(6319..6338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7132..7720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7621..7640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7874..7891
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8008..8029
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8044..8049,8050..8067,8068..8074)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8082..8098
                     /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     8106..8122
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8143..8161
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   8310..8344
                     /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 gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
      541 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc gcggccgcgg
     1741 taccggcgcg ccggaagttc ctatactttc tagagaatag gaacttcata tcgccaccat
     1801 ggccagctcc gaggatgtca tcaaagagtt tatgagattt aaggtcaaga tggagggaag
     1861 cgtcaacgga cacgagttcg agattgaggg agaaggagaa ggccggcctt acgagggcac
     1921 acaaaccgct aagctcaagg tcacaaaagg aggacccctc cccttctcct gggatattct
     1981 gagccctcag ttccagtacg gaagcaaagc ctatgttaaa caccctgccg acatccctga
     2041 ctatctgaag ctctccttcc ctgaaggctt caagtgggag agattcatga acttcgagga
     2101 cggaggcgtg gtgacagtca cacaagatag caccctccag gacggagagt ttatttataa
     2161 ggtgaaactc agaggaacca acttcccctc cgatggccct gtcatgcaaa aaaaaacaat
     2221 gggatgggaa gcctccaccg agagaatgta tcctgaggat ggcgctctga aaggcgaaat
     2281 taaaatgaga ctgaaactca aagacggagg acactacgat gccgaggtca aaacaaccta
     2341 caaggccaag aaacaagtgc agctgcctgg cgcctacatg actgatatta aactcgacat
     2401 tatcagccat aatggggact acaccatcgt ggaacaatat gagagagctg agggcagaca
     2461 tagcacaggc gctggagagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa
     2521 tcccggccct atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     2581 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     2641 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     2701 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     2761 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     2821 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     2881 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaaa atcaacctct
     2941 ggacgaaatt ccgcggttat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3001 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcag ttaacttgtt
     3061 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3121 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3181 ctggaattga ctcaagaagt tcctatactt tctagagaat aggaacttca tgatgtggcg
     3241 cgccggatcc tgcaggttat cacaagtttg tacaaaaaag caggctttaa aggaaccgcc
     3301 ttttgctggc cttttgctca catgtgaggg cctatttccc atgattcctt catatttgca
     3361 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
     3421 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
     3481 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
     3541 ggctttatat atcttgtgga aaggacgaaa caccgttggg gagacgagct ttcaagtttt
     3601 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     3661 cgagtcggtg cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     3721 ttttagcgcg tgcgccaatt ctgcagacaa atggctctag agaccggcgc cgctacaggg
     3781 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     3841 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     3901 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     3961 ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcctccagc ttttgttccc
     4021 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4081 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4141 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4201 accggtggtc tctctagagg tacccgttac ataacttacg gtaaatggcc cgatctagac
     4261 ccagctttct tgtacaaagt ggtgataact ctagagaatt cactcctcag gtgcaggctg
     4321 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     4381 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     4441 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     4501 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     4561 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     4621 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     4681 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     4741 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     4801 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttgga tccctcgagt
     4861 taattaacga gagcataata ttgatatgtg ccaaagttgt ttctgactga ctaataagta
     4921 taatttgttt ctattatgta taggttaagc taattactta ttttataata caacatgact
     4981 gtttttaaag tacaaaataa gtttattttt gtaaaagaga gaatgtttaa aagttttgtt
     5041 actttataga agaaattttg agtttttgtt tttttttaat aaataaataa acataaataa
     5101 attgtttgtt gaatttatta ttagtatgta agtgtaaata taataaaact taatatctat
     5161 tcaaattaat aaataaacct cgatatacag accgataaaa cacatgcgtc aattttacgc
     5221 atgattatct ttaacgtacg tcacaatatg attatctttc tagggttaaa taatagtttc
     5281 taattttttt attattcagc ctgctgtcgt gaataccgag ctccaattcg ccctatagtg
     5341 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     5401 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     5461 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     5521 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5581 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5641 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5701 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5761 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5821 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5881 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5941 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     6001 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6061 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6121 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6181 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6241 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6301 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6361 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6421 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6481 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6541 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6601 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6661 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6721 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6781 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6841 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6901 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6961 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     7021 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7081 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7201 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7261 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7741 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7801 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7861 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     7921 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     7981 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     8041 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     8101 tcacacagga aacagctatg accatgatta cgccaagctc gaaattaacc ctcactaaag
     8161 ggaacaaaag ctggtacctc gcgcgacttg gtttgccatt ctttagcgcg cgtcgcgtca
     8221 cacagcttgg ccacaatgtg gtttttgtca aacgaagatt ctatgacgtg tttaaagttt
     8281 aggtcgagta aagcgcaaat cttttttaac cctagaaaga tagtctgcgt aaaattgacg
     8341 catgcattct tgaaatattg ctctctcttt ctaaatagcg cgaatccgtc gctgtgcatt
     8401 taggacatct cagtcgccgc ttggagctcc cgtgaggcgt gcttgtcaat gcggtaagtg
     8461 tcactgattt tgaactataa cgaccgcgtg agtcaaaatg acgcatgatt atcttttacg
     8521 tgacttttaa gatttaactc atacgataat tatattgtta tttcatgttc tacttacgtg
     8581 ataacttatt atatatatat tttcttgtta tagatatcgt gactaatata taataaaatg
     8641 ggtagttctt tagacgatga gcatatcctc tctgctcttc tgcaaagcga tgacgagctt
     8701 gttggctagc ggtggcggcc tc
//
