LOCUS       sequence-271433-        9134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac vector for expression of 3xgRNA targeting TCF4 for CRISPRi;
            mRFP-T2A-BlasticidinR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9134)
  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 9134)
  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: pPN441
FEATURES             Location/Qualifiers
     source          1..9134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2494..2515
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2530..2535,2536..2553,2554..2560)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2568..2584
                     /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     2573..2595
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2592..2608
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2592..2608
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2629..2647
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2796..2830
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        3209..3588
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3590..3867
                     /label=chicken beta-actin promoter
     intron          3868..4884
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4808..4830
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4892..4911
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(4962..4995)
                     /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      5001..5010
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5685..5738
                     /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             5739..6137
                     /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(6207..6227)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    6261..6382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6298..6317)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6352..6371
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(6404..6437)
                     /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    6470..6494
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        6537..6785
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        6807..6882
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6980..7228
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7250..7325
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7423..7671
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7693..7768
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(7879..7903)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7999..8018)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8064..8119
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8118..8137)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   8823..8885
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8951..8970)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8952..8970)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8977..8994)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8977..8993)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8986..9008)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     2401 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     2461 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     2521 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     2581 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctcgaaa ttaaccctca
     2641 ctaaagggaa caaaagctgg tacctcgcgc gacttggttt gccattcttt agcgcgcgtc
     2701 gcgtcacaca gcttggccac aatgtggttt ttgtcaaacg aagattctat gacgtgttta
     2761 aagtttaggt cgagtaaagc gcaaatcttt tttaacccta gaaagatagt ctgcgtaaaa
     2821 ttgacgcatg cattcttgaa atattgctct ctctttctaa atagcgcgaa tccgtcgctg
     2881 tgcatttagg acatctcagt cgccgcttgg agctcccgtg aggcgtgctt gtcaatgcgg
     2941 taagtgtcac tgattttgaa ctataacgac cgcgtgagtc aaaatgacgc atgattatct
     3001 tttacgtgac ttttaagatt taactcatac gataattata ttgttatttc atgttctact
     3061 tacgtgataa cttattatat atatattttc ttgttataga tatcgtgact aatatataat
     3121 aaaatgggta gttctttaga cgatgagcat atcctctctg ctcttctgca aagcgatgac
     3181 gagcttgttg gctagcggtg gcggcctcga cattgattat tgactagtta ttaatagtaa
     3241 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     3301 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     3361 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     3421 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     3481 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     3541 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc
     3601 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
     3661 tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca
     3721 ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc
     3781 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
     3841 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
     3901 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     3961 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
     4021 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
     4081 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
     4141 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
     4201 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
     4261 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
     4321 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
     4381 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
     4441 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
     4501 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
     4561 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
     4621 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
     4681 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
     4741 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
     4801 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
     4861 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     4921 cattttggca aagaattcgc ggccgcggta ccggcgcgcc ggaagttcct atactttcta
     4981 gagaatagga acttcatatc gccaccatgg ccagctccga ggatgtcatc aaagagttta
     5041 tgagatttaa ggtcaagatg gagggaagcg tcaacggaca cgagttcgag attgagggag
     5101 aaggagaagg ccggccttac gagggcacac aaaccgctaa gctcaaggtc acaaaaggag
     5161 gacccctccc cttctcctgg gatattctga gccctcagtt ccagtacgga agcaaagcct
     5221 atgttaaaca ccctgccgac atccctgact atctgaagct ctccttccct gaaggcttca
     5281 agtgggagag attcatgaac ttcgaggacg gaggcgtggt gacagtcaca caagatagca
     5341 ccctccagga cggagagttt atttataagg tgaaactcag aggaaccaac ttcccctccg
     5401 atggccctgt catgcaaaaa aaaacaatgg gatgggaagc ctccaccgag agaatgtatc
     5461 ctgaggatgg cgctctgaaa ggcgaaatta aaatgagact gaaactcaaa gacggaggac
     5521 actacgatgc cgaggtcaaa acaacctaca aggccaagaa acaagtgcag ctgcctggcg
     5581 cctacatgac tgatattaaa ctcgacatta tcagccataa tggggactac accatcgtgg
     5641 aacaatatga gagagctgag ggcagacata gcacaggcgc tggagagggc agaggaagtc
     5701 ttctaacatg cggtgacgtg gaggagaatc ccggccctat ggccaagcct ttgtctcaag
     5761 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     5821 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     5881 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     5941 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     6001 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     6061 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     6121 atgtgtggga gggctaaaat caacctctgg acgaaattcc gcggttatta caaaatttgt
     6181 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     6241 ttaatgcctt tgtatcagtt aacttgttta ttgcagctta taatggttac aaataaagca
     6301 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6361 ccaaactcat caatgtatct tatcatgtct ggaattgact caagaagttc ctatactttc
     6421 tagagaatag gaacttcatg atgtggcgcg ccggatcctg caggttatca caagtttgta
     6481 caaaaaagca ggctttaaag gaaccaattc agtcgactgg atccggtact acatgtgagg
     6541 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
     6601 ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
     6661 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
     6721 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
     6781 acaccgtaaa cttgttccaa gtttaggttt tagagctaga aatagcaagt taaaataagg
     6841 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttgt tttagagcta
     6901 gaaatagcaa gttaaaataa ggctagtccg tttttagcgc gtgcgccaat tctgcagaca
     6961 aatggctcta gaggcatgtg agggcctatt tcccatgatt ccttcatatt tgcatatacg
     7021 atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa agatattagt
     7081 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     7141 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     7201 atatatcttg tggaaaggac gaaacaccgc atcaccatgg actcccccgg ttttagagct
     7261 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     7321 ggtgcttttt tgttttagag ctagaaatag caagttaaaa taaggctagt ccgtttttag
     7381 cgcgtgcgcc aattctgcag acaaatggct ctagaggcat gtgagggcct atttcccatg
     7441 attccttcat atttgcatat acgatacaag gctgttagag agataattgg aattaatttg
     7501 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
     7561 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
     7621 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cgtttcctca
     7681 aacaattctt gtgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     7741 acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta gagctagaaa tagcaagtta
     7801 aaataaggct agtccgtttt tagcgcgtgc gccaattctg cagacaaatg gctctagagg
     7861 taccctcgag atatctagac ccagctttct tgtacaaagt ggtgataact ctagagaatt
     7921 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     7981 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     8041 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     8101 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     8161 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     8221 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     8281 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     8341 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     8401 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     8461 caagcttgga tccctcgagt taattaacga gagcataata ttgatatgtg ccaaagttgt
     8521 ttctgactga ctaataagta taatttgttt ctattatgta taggttaagc taattactta
     8581 ttttataata caacatgact gtttttaaag tacaaaataa gtttattttt gtaaaagaga
     8641 gaatgtttaa aagttttgtt actttataga agaaattttg agtttttgtt tttttttaat
     8701 aaataaataa acataaataa attgtttgtt gaatttatta ttagtatgta agtgtaaata
     8761 taataaaact taatatctat tcaaattaat aaataaacct cgatatacag accgataaaa
     8821 cacatgcgtc aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc
     8881 tagggttaaa taatagtttc taattttttt attattcagc ctgctgtcgt gaataccgag
     8941 ctccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     9001 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     9061 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     9121 gaatggcgaa tggg
//
