LOCUS       sequence-474706-        8847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Perturb Seq sgRNA vector with PiggyBac backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8847)
  AUTHORS   Sivakumar S, Wang Y, Goetsch SC, Pandit V, Wang L, Zhao H, 
            Sundarrajan A, Armendariz D, Takeuchi C, Nzima M, Chen WC, Dederich 
            AE, El Hayek L, Gao T, Ghazawi R, Gogate A, Kaur K, Kim HB, McCoy 
            MK, Niederstrasser H, Oura S, Pinzon-Arteaga CA, Sanghvi M, Schmitz 
            DA, Yu L, Zhang Y, Zhou Q, Kraus WL, Xu L, Wu J, Posner BA, Chahrour
            MH, Hon GC, Munshi NV
  TITLE     Benchmarking and optimizing Perturb-seq in differentiating human 
            pluripotent stem cells.
  JOURNAL   bioRxiv [Preprint]. 2025 Jan 23:2025.01.21.633969. doi: 
            10.1101/2025.01.21.633969.
  PUBMED    39896670
REFERENCE   2  (bases 1 to 8847)
  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: "bioRxiv 
            [Preprint]. 2025 Jan 23:2025.01.21.633969. doi: 
            10.1101/2025.01.21.633969."
COMMENT     Sequence Label: PB-U6-sgRNA-BFP-Puro
FEATURES             Location/Qualifiers
     source          1..8847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   126..188
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    207..256
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     primer_bind     complement(289..309)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(289..307)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(328..344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(328..344)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(341..363)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    352..368
                     /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(376..382,383..400,401..406))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    421..442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(559..576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(730..1318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(810..829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1489..2280,2281..2349))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIDEIGAS
                     LIKHW"
     primer_bind     2112..2131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2350..2454)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2480..2935)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2617..2638)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2829..2848
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3062..3084
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3076..3093
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3077..3093
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3103..3122
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3103..3121
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3150..3183
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     repeat_region   3202..3236
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     CDS             4422..4466
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        4644..4892
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4644..4664
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4815..4834
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    5080..5197
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5336..6514
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5567..6505
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6462..6482
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7256..7312
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7313..7909
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7806..7826
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7947..8535
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8000..8020)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8418..8429)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    8568..8792
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat gtaagtgtaa
       61 atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata cagaccgata
      121 aaacacatgc gtcaatttta cgcatgatta tctttaacgt acgtcacaat atgattatct
      181 ttctagggtt aatctagtat acgcgttcgg ccggcttgtc gacgacggcg gtctccgtcg
      241 tcaggatcat ccgggcatgc ggccgcttaa ttaatccagc ttttgttccc tttagtgagg
      301 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      361 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      421 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      481 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      541 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      601 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      661 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      721 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      781 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      841 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      901 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      961 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1021 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1081 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1141 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1201 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1261 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1321 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1381 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1441 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1501 aatcagtgag gcacctatct catcgatctg tctatttcgt tcatccatag ttgcctgact
     1561 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1621 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1681 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1741 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1801 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1861 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1921 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1981 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2041 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2101 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2161 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2221 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2281 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2341 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2401 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2461 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2521 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     2581 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2641 ttaaagaacg tggattccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2701 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2761 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2821 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     2881 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     2941 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     3001 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     3061 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     3121 ggcgaattgg ggcgcgcccc gcggtgcggg tgccagggcg tgcccttggg ctccccgggc
     3181 gcgtactcca ttctagatta accctagaaa gatagtctgc gtaaaattga cgcatgcatt
     3241 cttgaaatat tgctctctct ttctaaatag cgcgaatccg tcgctgtgca tttaggacat
     3301 ctcagtcgcc gcttggagct cccgtgaggc gtgcttgtca atgcggtaag tgtcactgat
     3361 tttgaactat aacgaccgcg tgagtcaaaa tgacgcatga ttatctttta cgtgactttt
     3421 aagatttaac tcatacgata attatattgt tatttcatgt tctacttacg tgataactta
     3481 ttatatatat attttcttgt tatagatatc aactagaatg gggaggtggt ccctgcagtt
     3541 acgccaatga taacccccgc cagaaaaatc ttagtagcct tccctttttg ttttccgtgc
     3601 cccaactcgg cggattgact cggccccttc cggaaacacc cgaatcaact tctagtcaaa
     3661 ttattgttca cgccgcaatg acccacccct ggcccgcgtc tgtggaactg acccctggtg
     3721 tacaggagag ttcgctgctg aaagtggtcc caaaggggta ctagttttta agctcccaac
     3781 tccccctccc ccagcgtctg gaggattcca caccctcgca ccgcaggggc gaggaagtgg
     3841 gcggagtccg gttttggcgc cagccgctga ggctgccaag cagaaaagcc accgctgagg
     3901 agactccggt cactgtcctc gccccgcctc ccccttccct ccccttgggg accaccgggc
     3961 gccacgccgc gaacggtaag tgccgcggtc gtcggcgcct ccgccctccc cctagggccc
     4021 caattcccag cgggcgcggc gcgcggcccc tccccccgcc gggcgcgcgc ccgctgcccc
     4081 gcccttcgtg gccgcccggc gtgggcggtg ccacccctcc ccccggcggc cccgcgcgca
     4141 gctcccggct ccctccccct tcggatgtgg cttgagctgt aggcgcggag ggccctgcaa
     4201 agatggataa agttttaaac agagaggaat ctttgcagct aatggacctt ctaggtcttg
     4261 aaaggagtgg gaattctgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     4321 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     4381 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     4441 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     4501 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     4561 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     4621 tcccaacccc gaggggaccc agagagggcc tatttcccat gattccttca tatttgcata
     4681 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat
     4741 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     4801 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     4861 ctttatatat cttgtggaaa ggacgaaaca ccggagacgg gataccgtct ctgtttaaga
     4921 gctaagctgg aaacagcata gcaagtttaa ataaggctag tccgttatca acttggccgc
     4981 tttaaggccg gtcctagcaa ggccaagtgg caccgagtcg gtgctttttt aagcttggcg
     5041 taactagatc ttgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     5101 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     5161 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     5221 agatccactt tggcgccggc tcgagggggc ccgggtgcaa agatggataa agttttaaac
     5281 agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc
     5341 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     5401 ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     5461 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     5521 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg
     5581 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc
     5641 cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
     5701 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
     5761 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
     5821 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
     5881 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
     5941 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
     6001 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
     6061 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     6121 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     6181 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     6241 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     6301 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     6361 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     6421 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     6481 tcaaagtttt tttcttccat ttcaggtgtc gtgagagctc ggtaccgcgg gcccgggatc
     6541 caccatggtc tccaagggtg aggaactgat caaagaaaat atgcatatga agctgtacat
     6601 ggaggggacc gtggataacc accacttcaa gtgtactagt gagggcgaag gcaaaccata
     6661 tgaaggtacg cagacaatga gaatcaaggt tgttgaaggc ggaccactgc cttttgcctt
     6721 tgacattctt gctaccagtt tcctttacgg ttcaaaaacc tttataaacc acacacaggg
     6781 gatccctgat ttcttcaagc agagctttcc tgaagggttt acttgggaga gagtgactac
     6841 gtatgaggac ggcggtgtcc tgacagccac ccaggataca tcacttcagg acggctgtct
     6901 tatttataat gtgaagatca gaggcgtgaa tttcacttcc aacggccccg tgatgcagaa
     6961 aaaaacactc ggctgggagg cctttacgga aacactttat ccagctgatg gcggtctgga
     7021 gggaagaaac gacatggctt tgaagctggt ggggggaagt catctgattg ccaacgccaa
     7081 gacaacttac cggtctaaaa aaccggccaa gaatctgaaa atgcctggcg tctactatgt
     7141 cgactatcgc ctggaacgga tcaaggaggc taataacgag acttacgtgg agcagcatga
     7201 ggtcgctgtc gcaagatact gtgatcttcc tagcaaactt ggccataagc tgaacgcaac
     7261 aaacttctct ctgctgaaac aagccggaga tgtcgaagag aatcctggac cgaccgagta
     7321 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     7381 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     7441 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     7501 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     7561 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     7621 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     7681 cctggccacc gtcggagtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     7741 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     7801 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     7861 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag gatccaatca
     7921 acctctggat tacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     7981 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     8041 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     8101 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     8161 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     8221 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     8281 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     8341 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     8401 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     8461 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     8521 gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg tgccttctag
     8581 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     8641 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     8701 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     8761 caggcatgct ggggatgcgg tgggctctat ggctagttaa aagttttgtt actttataga
     8821 agaaattttg agtttttgtt ttttttt
//
