LOCUS       sequence-375206-        9232 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid vector for in vivo immune checkpoint screening library (for 
            checkpoint library).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9232)
  AUTHORS   Lu Z, Ni K, Wang Y, Zhou Y, Li Y, Yan J, Song Q, Liu M, Xu Y, Yu Z, 
            Guo T, Ma L
  TITLE     An in-library ligation strategy and its application in CRISPR/Cas9 
            screening of high-order gRNA combinations.
  JOURNAL   Nucleic Acids Res. 2022 Jun 24;50(11):6575-6586. doi: 
            10.1093/nar/gkac458.
  PUBMED    35670669
REFERENCE   2  (bases 1 to 9232)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2022-06-24- 24"; volume: "50"; issue: 
            "11"; pages: "6575-6586"
COMMENT     Sequence Label: Immune checkpoint (library 2) Backbone
FEATURES             Location/Qualifiers
     source          1..9232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..42)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(702..1493,1494..1562))
                     /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     1325..1344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1563..1667)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1693..2148)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1830..1851)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2042..2061
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2275..2297
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2289..2306
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2290..2306
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2316..2335
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2316..2334
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2355..2490
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2530..2651
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2541..2560)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2595..2614
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2729..2962
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3034..3622
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3140..3151
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3549..3569
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3644..4339)
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     CDS             complement(4340..4393)
                     /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             complement(4394..4990)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4474..4494)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4971..4990
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5003..6181)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5012..5950
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5035..5055)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    6320..6437
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6493..6741
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6493..6513
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6664..6683
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             complement(7172..7216)
                     /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"
     misc_feature    7401..7634
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    8127..8252
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     promoter        complement(8480..8706)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(8734..8754)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8734..8752)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8773..8789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8773..8789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8786..8808)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8797..8813
                     /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(8821..8827,8828..8845,8846..8851))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8866..8887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9004..9021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(9175..9232,1..531))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
       61 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      121 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      181 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      241 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      301 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      361 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      601 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      661 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      721 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
      781 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
      841 cgggacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
      901 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
      961 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1021 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1081 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1141 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1201 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1261 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1321 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1381 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1441 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1501 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1561 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1621 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1681 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     1741 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     1801 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     1861 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatcgc ccactacgtg
     1921 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     1981 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2041 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2101 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     2161 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     2221 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     2281 cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat
     2341 tgggtacgtc cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     2401 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     2461 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga tagctagagc
     2521 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2581 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2641 ataaacaagt tcctctcact ctctgatatt catttctttg caagttataa atactgaata
     2701 ataagatgac atgaactact atacgtactg ctagagattt tccacactga ctaaaagggt
     2761 ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact
     2821 caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag
     2881 gctcagatct ggtctaacca gagagagcca gtacaagcaa aaagcagatc ttgtcatcgt
     2941 tgggagtgaa ttagcccttc cagtcccccc ttttctttta aaaagtggct aagatctaca
     3001 gctgccttgt aagtcattgg tcttaaagtc gacgcgggga ggcggcccaa agggagatcc
     3061 gactcgtctg agggcgaagg cgaacacgcg gaagaggccg cagagccggc agcaggccgc
     3121 gggaaggaag gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca
     3181 gaatccaggt ggcaacacag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca
     3241 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
     3301 cgttgagttc cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac
     3361 aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc
     3421 cacgttgcct gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca
     3481 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
     3541 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
     3601 ttttgtaatc cagaggttga ttgtcgactt aacgcgtgga tccttatctg tgccccagtt
     3661 tgctagggag gtcgcagtat ctggccacag ccacctcgtg ctgctcgacg taggtttctt
     3721 tgtcggcctc cttgattctt tccagccttc tgtccacata gtagacgccg ggcatcttga
     3781 ggttcttagc gggtttcttg gatctgtatg tggttttcaa gttgcagatc aggtggcccc
     3841 cgcccacgag cttcagggcc atgtcggctc tgccttccag gccgccgtca gcggggtaca
     3901 gggtttcggt ggaggcctcc cagccgagtg ttttcttctg catcacaggg ccgttggatg
     3961 ggaagttcac ccctctgatc ttgacgttgt agatgaggca gccgtcctgg aggctggtgt
     4021 cctgggtagc ggtcagcacg cccccgtcct cgtatgtggt gactctctcc catgtgaagc
     4081 cctcggggaa ggactgctta aagaagtcgg ggatgccctg ggtgtggttg atgaaggttt
     4141 tgctgccgta catgaagctg gtagccagga tgtcgaaggc gaaggggaga gggccgccct
     4201 cgaccgcctt gattctcatg gtctgggtgc cctcgtaggg cttgccttcg ccctcggatg
     4261 tgcacttgaa gtggtggttg ttcacggtgc cctccatgta cagcttcatg tgcatgttct
     4321 ccttaatcag ctcgctcacg gggccgggat tttcctccac gtccccgcat gttagcagac
     4381 ttcccctgcc ctcggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca
     4441 cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgcggggcg
     4501 cggaggtttc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca
     4561 cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gactccgacg gtggccagga
     4621 accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg
     4681 ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg
     4741 cttcgacgct ctccggcgtg gtccagaccg ccacggcggc gccgtcgtcc gcgacccaca
     4801 ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct
     4861 cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg
     4921 cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt
     4981 actcggtcat ggtggccgta cgtcacgaca cctgaaatgg aagaaaaaaa ctttgaacca
     5041 ctgtctgagg cttgagaatg aaccaagatc caaactcaaa aagggcaaat tccaaggaga
     5101 attacatcaa gtgccaagct ggcctaactt cagtctccac ccactcagtg tggggaaact
     5161 ccaacgcata aaacccctcc ccccaaccta aagacgacgt actccaaaag ctcgagaact
     5221 aatcgaggtg cctggacggc gcccggtact ccgtggagtc acatgaagcg acggctgagg
     5281 acggaaaggc ccttttcctt tgtgtgggtg actcacccgc ccgctctccc gagcgccgcg
     5341 tcctccattt tgagctccct gcagcagggc cgggaagcgg ccatctttcc gctcacgcaa
     5401 ctggtgccga ccgggccagc cttgccgccc agggcggggc gatacacggc ggcgcgaggc
     5461 caggcaccag agcaggccgg ccagcttgag actacccccg tccgattctc ggtggccgcg
     5521 ctcgcaggcc ccgcctcgcc gaacatgtgc gctgggacgc acgggccccg tcgccgcccg
     5581 cggccccaaa aaccgaaata ccagtgtgca gatcttggcc cgcatttaca agactatctt
     5641 gccagaaaaa aagcgtcgca gcaggtcatc aaaaatttta aatggctaga gacttatcga
     5701 aagcagcgag acaggcgcga aggtgccacc agattcgcac gcggcggccc cagcgcccag
     5761 gccaggcctc aactcaagca cgaggcgaag gggctcctta agcgcaaggc ctcgaactct
     5821 cccacccact tccaacccga agctcgggat caagaatcac gtactgcagc caggtggaag
     5881 taattcaagg cacgcaaggg ccataacccg taaagaggcc aggcccgcgg gaaccacaca
     5941 cggcacttac ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta
     6001 ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat
     6061 cactttccca gtttaccccg cgccaccttc tctaggcacc ggatcaattg ccgacccctc
     6121 cccccaactt ctcggggact gtgggccatg tgcgctctgc ccactgacgg gcaccggagc
     6181 caattcccac tcctttcaag acctagaagg tccattagct gcaaagattc ctctctgttt
     6241 aaaactttat ccatctttgc acccgggccc cctcgagccg gcgccaaagt ggatctctgc
     6301 tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt aattctttag
     6361 tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac cccccaatcc
     6421 ccccttttct tttaaaattg tggatgaata ctgccatttg tctcaagatc tagttacgcc
     6481 aagcttggta ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag
     6541 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
     6601 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     6661 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     6721 ttgtggaaag gacgaaacac cggagacggt tgtaaagata tctgagcaca caaaatatct
     6781 agaatccatg ctaaaatatt gttcctgttc cattaactgc gctaaaaata aacgtctctg
     6841 tttgagagct atgctggagc ccgaacagca tagcaagttc aaataaggct agtccgttat
     6901 caacttggcc cgaaaaaaaa gaaaaaaaga aaaaaagaaa aacgggccaa gtggcaccga
     6961 gtcggtgctt tttttcaccg tggaatgcga gaattctggg tcccctcggg gttgggaggt
     7021 gggtctgaaa cgataatggt gaatatccct gcctaactct attcactata gaaagtacag
     7081 caaaaactat tcttaaacct accaagcctc ctactatcat tatgaataat tttatatacc
     7141 acagccaatt tgttatgtta aaccaattcc acaaacttgc ccatttatct aattccaata
     7201 attcttgttc attcttttct tgctggtttt gcgattcttc aattaaggag tgtattaagc
     7261 ttgtgtaatt gttaatttct ctgtcccact ccatccaggt cgtgtgattc caaatctgtt
     7321 ccagagattt attactccaa ctagcattcc aaggcacagc agtggtgcaa atgagttttc
     7381 cagagcaacc ccaaatcccc aggagctgtt gatcctttag gtatctttcc acagccagga
     7441 ttcttgcctg gagctgcttg atgccccaga ctgtgagttg caacagatgc tgttgcgcct
     7501 caatagccct cagcaaattg ttctgctgct gcactatacc agacaataat tgtctggcct
     7561 gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc tgctgctccc aagaacccaa
     7621 ggaacaaagc tcctattccc actgctcttt tttctctctg caccactctt ctctttgcct
     7681 tggtgggtgc tactcctaat ggttcaattt ttactacttt atatttatat aattcacttc
     7741 tccaattgtc cctcatatct cctcctccag gtctgaagat cagcggccgc ttgctgtgcg
     7801 gtggtcttac ttttgttttg ctcttgctct atcttgtcta aagcttcctt ggtgtctttt
     7861 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     7921 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     7981 gatgtttcta acaggccagg attaactgcg aatcgttcta gctccctgct tgcccatact
     8041 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccaacgc
     8101 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     8161 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     8221 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     8281 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     8341 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     8401 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     8461 ggtctaacca gagagagccg tttatgtatc gagctaggca cttaaataca atatctctgc
     8521 aatgcggcaa ttcagtggtt cgtccaatcc atgtcagacc cgtctgttgc cttcctaata
     8581 aggcacgatc gtaccacctt acttccacca atcggcatgc acggtgcttt ttctctcctt
     8641 gtaaggcatg ttgctaactc atcgttacca tgttgcaaga ctacaagagt attgcataag
     8701 actacattaa gcttgcagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     8761 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     8821 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     8881 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     8941 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcatccgc
     9001 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     9061 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     9121 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     9181 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga gg
//
