LOCUS       sequence-474710-       12826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Perturb Seq landing pad with Pa01 recombinase at the AAVS1 loci.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12826)
  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 12826)
  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: AAVS1-SA-CAG-Pa01-BFP
FEATURES             Location/Qualifiers
     source          1..12826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    22..858
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(935..952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1106..1694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1186..1205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1865..2656,2657..2725))
                     /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     2488..2507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2726..2830)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2898..2916
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2954..2976)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3076..3095
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    3245..4048
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    complement(4078..4111)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4112..4137
                     /label=SA
                     /note="splice acceptor site"
     CDS             4161..4214
                     /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             4209..5009
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPRIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4269..4288)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4879..4898
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5014..5238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        5274..5485
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5498..5766
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      5804..5813
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5955..5973)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6223..6242
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(6565..6582)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6571..6795
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    8393..8981
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8446..8466)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8864..8875)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     enhancer        9205..9508
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9510..9786
                     /label=chicken beta-actin promoter
     intron          9787..10803
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     10727..10749
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     10811..10830
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             12476..12523
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    12570..12691
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gggtcgacag tactaagctt tactagggac aggattggtg acagaaaagc cccatcctta
       61 ggcctcctcc ttcctagtct cctgatattg ggtctaaccc ccacctcctg ttaggcagat
      121 tccttatctg gtgacacacc cccatttcct ggagccatct ctctccttgc cagaacctct
      181 aaggtttgct tacgatggag ccagagagga tcctgggagg gagagcttgg cagggggtgg
      241 gagggaaggg ggggatgcgt gacctgcccg gttctcagtg gccaccctgc gctaccctct
      301 cccagaacct gagctgctct gacgcggctg tctggtgcgt ttcactgatc ctggtgctgc
      361 agcttcctta cacttcccaa gaggagaagc agtttggaaa aacaaaatca gaataagttg
      421 gtcctgagtt ctaactttgg ctcttcacct ttctagtccc caatttatat tgttcctccg
      481 tgcgtcagtt ttacctgtga gataaggcca gtagccagcc ccgtcctggc agggctgtgg
      541 tgaggagggg ggtgtccgtg tggaaaactc cctttgtgag aatggtgcgt cctaggtgtt
      601 caccaggtcg tggccgcctc tactcccttt ctctttctcc atccttcttt ccttaaagag
      661 tccccagtgc tatctgggac atattcctcc gcccagagca gggtcccgct tccctaaggc
      721 cctgctctgg gcttctgggt ttgagtcctt ggcaagccca ggagaggcgc tcaggcttcc
      781 ctgtccccct tcctcgtcca ccatctcatg cccctggctc tcctgcccct tccctacagg
      841 ggttcctggc tctgctctaa gggcgaattc tgcattaatg aatcggccaa cgcgcgggga
      901 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      961 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1021 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtgggcggtg
     1501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1741 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1801 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1861 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1921 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1981 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2041 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2101 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2161 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2221 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2281 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2341 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2401 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2461 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2521 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2581 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2641 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2701 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2821 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     2881 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     2941 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     3001 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     3061 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     3121 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     3181 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagaattcgc
     3241 cctttgcttt ctctgaccag cattctctcc cctgggcctg tgccgctttc tgtctgcagc
     3301 ttgtggcctg ggtcacctct acggctggcc cagatccttc cctgccgcct ccttcaggtt
     3361 ccgtcttcct ccactccctc ttccccttgc tctctgctgt gttgctgccc aaggatgctc
     3421 tttccggagc acttccttct cggcgctgca ccacgtgatg tcctctgagc ggatcctccc
     3481 cgtgtctggg tcctctccgg gcatctctcc tccctcaccc aaccccatgc cgtcttcact
     3541 cgctgggttc ccttttcctt ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg
     3601 gccttctccg acggatgtct cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat
     3661 caccgttttt ctggacaacc ccaaagtacc ccgtctccct ggctttagcc acctctccat
     3721 cctcttgctt tctttgcctg gacaccccgt tctcctgtgg attcgggtca cctctcactc
     3781 ctttcatttg ggcagctccc ctacccccct tacctctcta gtctgtgcta gctcttccag
     3841 ccccctgtca tggcatcttc caggggtccg agagctcagc tagtcttctt cctccaaccc
     3901 gggcccctat gtccacttca ggacagcatg tttgctgcct ccagggatcc tgtgtccccg
     3961 agctgggacc accttatatt cccagggccg gttaatgtgg ctctggttct gggtactttt
     4021 atctgtcccc tccaccccac agtggggcaa gcttcttacc cttctcccag cggccgcata
     4081 acttcgtata gcatacatta tacgaagtta tctgacctct tctcttcctc ccacagggcc
     4141 tcgagagatc tggcagcgga gagggcagag gaagtcttct aacatgcggt gacgtggagg
     4201 agaatcccgg ccctaggatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4261 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4321 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4381 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4441 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4501 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4561 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4621 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4681 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4741 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4801 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4861 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4921 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4981 tctatcgcct tcttgacgag ttcttctaaa aacctgtgcc ttctagttgc cagccatctg
     5041 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     5101 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     5161 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     5221 atgcggtggg ctctatggat gcatgatctg cgatcgctcc ggtgcccgtc agtgggcaga
     5281 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
     5341 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     5401 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     5461 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
     5521 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     5581 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     5641 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     5701 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
     5761 cagatccaag ctgtgaccgg cgcctacgct agcgctaccg gtcgccacca tggtgagcaa
     5821 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     5881 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     5941 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     6001 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     6061 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     6121 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     6181 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     6241 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     6301 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     6361 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     6421 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     6481 ccgccactcc accggcggca tggacgagta aggtaccata ctcgtatagc atacattata
     6541 cgaagttata aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     6601 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     6661 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     6721 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     6781 cggtgggctc tatgggtctc gggaggtggt ccctgcagtt acgccaatga taacccccgc
     6841 cagaaaaatc ttagtagcct tccctttttg ttttccgtgc cccaactcgg cggattgact
     6901 cggccccttc cggaaacacc cgaatcaact tctagtcaaa ttattgttca cgccgcaatg
     6961 acccacccct ggcccgcgtc tgtggaactg acccctggtg tacaggagag ttcgctgctg
     7021 aaagtggtcc caaaggggta ctagttttta agctcccaac tccccctccc ccagcgtctg
     7081 gaggattcca caccctcgca ccgcaggggc gaggaagtgg gcggagtccg gttttggcgc
     7141 cagccgctga ggctgccaag cagaaaagcc accgctgagg agactccggt cactgtcctc
     7201 gccccgcctc ccccttccct ccccttgggg accaccgggc gccacgccgc gaacggtaag
     7261 tgccgcggtc gtcggcgcct ccgccctccc cctagggccc caattcccag cgggcgcggc
     7321 gcgcggcccc tccccccgcc gggcgcgcgc ccgctgcccc gcccttcgtg gccgcccggc
     7381 gtgggcggtg ccacccctcc ccccggcggc cccgcgcgca gctcccggct ccctccccct
     7441 tcggatgtgg cttgagcttt tgataccgcg ggcagagctc gtttagtccg gtttcccttc
     7501 gcacccgcac cgcggcttcg agaccgtgac ctacatgctc gaagggcgta tgcgccacga
     7561 agaccacctc ggcaatcgcg gcctgctcaa gcaccatggt ctccaagggt gaggaactga
     7621 tcaaagaaaa tatgcatatg aagctgtaca tggaggggac cgtggataac caccacttca
     7681 agtgtactag tgagggcgaa ggcaaaccat atgaaggtac gcagacaatg agaatcaagg
     7741 ttgttgaagg cggaccactg ccttttgcct ttgacattct tgctaccagt ttcctttacg
     7801 gttcaaaaac ctttataaac cacacacagg ggatccctga tttcttcaag cagagctttc
     7861 ctgaagggtt tacttgggag agagtgacta cgtatgagga cggcggtgtc ctgacagcca
     7921 cccaggatac atcacttcag gacggctgtc ttatttataa tgtgaagatc agaggcgtga
     7981 atttcacttc caacggcccc gtgatgcaga aaaaaacact cggctgggag gcctttacgg
     8041 aaacacttta tccagctgat ggcggtctgg agggaagaaa cgacatggct ttgaagctgg
     8101 tggggggaag tcatctgatt gccaacgcca agacaactta ccggtctaaa aaaccggcca
     8161 agaatctgaa aatgcctggc gtctactatg tcgactatcg cctggaacgg atcaaggagg
     8221 ctaataacga gacttacgtg gagcagcatg aggtcgctgt cgcaagatac tgtgatcttc
     8281 ctagcaaact tggccataag ctgaactaat aatagcaaac tggggcacaa gcccctcaca
     8341 gccggatatc aagcttatcg gtaagctcac ctattagcgg ctaaggggat ccaatcaacc
     8401 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     8461 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     8521 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     8581 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     8641 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     8701 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     8761 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     8821 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     8881 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     8941 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgatccagac atgataagat
     9001 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     9061 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa cggtaccaga
     9121 ggttaacaac aacctgcagg aatctagtta ttaatagtaa tcaattacgg ggtcattagt
     9181 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     9241 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     9301 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     9361 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     9421 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     9481 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     9541 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
     9601 tgcagcgatg ggggcggggg gggggggggg cgcgcgccag gsggggsggg gsggggsgag
     9661 ggggggggsg gggsgaggsg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
     9721 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
     9781 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
     9841 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
     9901 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
     9961 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
    10021 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
    10081 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
    10141 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
    10201 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
    10261 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
    10321 gggcgtggcg cggggctcgc cgtgccgtgc ggggggtggc ggcaggkggg ggtgccgggc
    10381 ggggcggggc cgccycgggc cggggrgggc tcgggggrgg ggcgcggcgg cccccggagc
    10441 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
    10501 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
    10561 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
    10621 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
    10681 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
    10741 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
    10801 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgatt
    10861 tgataccgcg ggcagagctc gtttagtcac catgggccct agcgccttca gctacgtgcg
    10921 ctttagcagc ggcaagcagg ccaagggcag ctctgaacac agacagagag ccatgctggg
    10981 acagtggctg gaacagcacc ctagcttcac actgagtgat ctgcggttcg aggacctggg
    11041 cagaagcgga ttcagcggcg agcacctgga tcacggcctg ggccagctgc tggctgctat
    11101 cgacagcggc gccatcaaga gcggagatgt gatcctggtc gaggccgtgg acagaatcgg
    11161 cagactggag cctctggaaa tgctgcctct gttctccaga atcgtgaagg ccggcgtgtc
    11221 cgttatcacc ctggaagatg gccacgttta cgaccggagc tccgtgaacg agacatctct
    11281 gtttctgctg gtggccaaaa tccagcaggc ccacgagtac tctaatagac tgtctcgaag
    11341 aatcaacgcc tcttacaccg ctcggagaga aaaagctaaa gcaggcctgg gcatcaagag
    11401 agagacacct gtgtggctga ccacagacgg ccaactggtg cctcacgtgg cccctcacat
    11461 cgcccaggct tttcaggact acgccgatgg cctgggagag agaaggattt gtagaaagct
    11521 gcgggagagc ggcctggaag aattctccaa gaccaacgcc accaccgtgc ggagatggct
    11581 gaagaaccgg accgccattg gatattggaa cgacatccct gatgtgtacc ctcatgtcgt
    11641 ggaccctgct ctgttctacc aggtgcagca gcgcctggac gcccctaagg tggacagagc
    11701 caagcccagc gctcactacc tgacaggcct cgtgaagtgc gccgtgtgcg gcagaaacta
    11761 caactacaaa cagcggaagc acaccgaccc cgctatgctg tgcaccagca gagccagact
    11821 ggccggggaa ggctgcagca atagcaagac ataccccgtg atagttctgg accaggtgcg
    11881 gaagctgacc agcctgccgt tcctgcaaca cgccatggaa tccgcctcat ctcaggccga
    11941 cccaagcagc cagagactgg ccgtgatcga cggcgaaatc ggcgagctgt ccagaaagat
    12001 cagcgaggcc acaaaggccc tgctggtgct gggcttcacc cctgagatcc aggagagcct
    12061 ggaacagctc aagaccgcca gagaagccct ggaggaagaa agagccaccc tgctgctgcc
    12121 tcaggccgag aagctgacaa cagctcagct ggaggccttc agcaacggcc tgctggacga
    12181 cgagcccatg aagctcaacc acgtgcttca gaccgccggc tacagcatgg tggtgcaccc
    12241 cgacggctct atcgacgtgg atggaaaaag attcgtgtac gagggcgcta gccggaaaga
    12301 gaaagtgtac aagctgagac tgatcggcga ggataagcag tggagcctgc ccatcctgac
    12361 cccacagatg gccacttaca agtccctgtt catggccgct gtgcggctgc ctggcgaccc
    12421 cagcgaggaa gagctgagac ggttcgagga agccaagcac agcgagcggg gtggtaaaag
    12481 gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aagtaataat tctagagtcg
    12541 gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
    12601 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
    12661 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
    12721 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
    12781 taaaatcgat aaggatccgt cgaccgggta ccagagtaat tcaacc
//
