LOCUS       sequence-374824-        9823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral sgRNA vector for CROP-seq with mU6 sgRNA promoter, CR1 
            constant region, and UCOE EF1alpha driving PURO-BFP marker 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9823)
  AUTHORS   Replogle JM, Bonnar JL, Pogson AN, Liem CR, Maier NK, Ding Y, 
            Russell BJ, Wang X, Leng K, Guna A, Norman TM, Pak RA, Ramos DM, 
            Ward ME, Gilbert LA, Kampmann M, Weissman JS, Jost M
  TITLE     Maximizing CRISPRi efficacy and accessibility with dual-sgRNA 
            libraries and optimal effectors.
  JOURNAL   eLife. 2022 Dec 28;11:e81856. doi: 10.7554/eLife.81856.
  PUBMED    36576240
REFERENCE   2  (bases 1 to 9823)
  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: "eLife."; 
            date: "2022-12-28"; volume: "11:e81856. doi"; pages: " 
            10.7554/eLife.81856"
COMMENT     Sequence Label: pJR106
FEATURES             Location/Qualifiers
     source          1..9823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    100..121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(136..141,142..159,160..166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    174..190
                     /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     179..201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     198..214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     198..214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     233..253
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        235..253
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        281..507
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             508..688
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    735..860
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1353..1586
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1771..1815
                     /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    1996..2113
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2855..4033
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3086..4024
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3981..4001
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4045..4641
                     /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(4045..4064)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4541..4561
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4651..4704
                     /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             4717..5415
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     misc_feature    5461..6049
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5514..5534)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5932..5943)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(6405..6438)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     6442..6462
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     LTR             6690..6870
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    6948..7069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6985..7004)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7039..7058
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7088..7223
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7243..7262)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7244..7262)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7272..7289)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7272..7288)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7281..7303)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7430..7885
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7517..7536)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7727..7748
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7911..8015
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8016..8084,8085..8876)
                     /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(8234..8253)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9047..9635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9536..9555
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9789..9806
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
       61 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      121 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      181 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
      241 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
      301 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
      361 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
      421 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
      481 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
      541 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      601 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
      661 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
      721 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
      781 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
      841 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
      901 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
      961 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1021 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1081 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1141 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1201 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     1261 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     1321 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     1381 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     1441 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     1501 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     1561 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     1621 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     1681 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     1741 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     1801 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     1861 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     1921 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agaatcttga gacaaatggc
     1981 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     2041 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     2101 aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc gccggccgag
     2161 tactaggatc cattaggcgg ccgccgggag gtggtccctg cagttacgcc aatgataacc
     2221 cccgccagaa aaatcttagt agccttccct ttttgttttc cgtgccccaa ctcggcggat
     2281 tgactcggcc ccttccggaa acacccgaat caacttctag tcaaattatt gttcacgccg
     2341 caatgaccca cccctggccc gcgtctgtgg aactgacccc tggtgtacag gagagttcgc
     2401 tgctgaaagt ggtcccaaag gggtactagt ttttaagctc ccaactcccc ctcccccagc
     2461 gtctggagga ttccacaccc tcgcaccgca ggggcgagga agtgggcgga gtccggtttt
     2521 ggcgccagcc gctgaggctg ccaagcagaa aagccaccgc tgaggagact ccggtcactg
     2581 tcctcgcccc gcctccccct tccctcccct tggggaccac cgggcgccac gccgcgaacg
     2641 gtaagtgccg cggtcgtcgg cgcctccgcc ctccccctag ggccccaatt cccagcgggc
     2701 gcggcgcgcg gcccctcccc ccgccgggcg cgcgcccgct gccccgccct tcgtggccgc
     2761 ccggcgtggg cggtgccacc cctccccccg gcggccccgc gcgcagctcc cggctccctc
     2821 ccccttcgga tgtggcttga gctgtaggcg cggagatgca tgtgcccgtc agtgggcaga
     2881 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
     2941 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     3001 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     3061 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
     3121 ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat
     3181 tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg
     3241 agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg
     3301 aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa
     3361 tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca
     3421 agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc
     3481 ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg
     3541 gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc
     3601 gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct
     3661 tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg
     3721 tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc
     3781 cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgcgctttt ggagtacgtc
     3841 gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga
     3901 gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga
     3961 gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt
     4021 tcaggtgtcg tgagctagcc caccatgacc gagtacaagc ccacggtgcg cctcgccacc
     4081 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     4141 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     4201 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     4261 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     4321 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     4381 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtgtcgccc
     4441 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     4501 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4561 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4621 atgacccgca agcccggtgc cggatcggga gagggcagag gaagtctgct aacatgcggt
     4681 gacgtcgagg agaatcctgg cccaccggtc gccaccagcg agctgattaa ggagaacatg
     4741 cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag
     4801 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc
     4861 cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc
     4921 atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca
     4981 tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     5041 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac
     5101 ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac cctgtacccc
     5161 gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat
     5221 ctgatcgcaa acatcaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg
     5281 cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc
     5341 tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg
     5401 cacaagctta attaagaatt cgtcgaggga cctaataact attaaacgcg ttaagtcgac
     5461 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     5521 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     5581 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     5641 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     5701 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5761 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     5821 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     5881 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     5941 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     6001 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     6061 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     6121 tggaagggct aattcactcc caacgaagac aagatttcac cattatcgtt tcagacccac
     6181 ctcccaaccc cgaggggacc cagatctaga gatccgacgc gccatctcta ggcccgcgcc
     6241 ggccccctcg cacggacttg tgggagaagc tcggctactc ccctgccccg gttaatttgc
     6301 atataatatt tcctagtaac tatagaggct taatgtgcga taaaagacag ataatctgtt
     6361 ctttttaata ctagctacat tttacatgat aggcttggat ttctataact tcgtatagca
     6421 tacattatac gaagttataa acagcacaaa aggaaactca ccctaactgt aaagtaattg
     6481 tgtgttttga gactataagt atcccttgga gaaccacctt gttggaccag gatgggcacc
     6541 acccgtttaa gagctaagct ggaaacagca tagcaagttt aaataaggct agtccgttat
     6601 caacttgaaa aagtggcacc gagtcggtgc tttttttctc gagaagcttg gcgtaactag
     6661 atcttgagac actgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6721 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6781 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6841 ccttttagtc agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta
     6901 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6961 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     7021 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     7081 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     7141 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     7201 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     7261 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     7321 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     7381 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     7441 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     7501 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     7561 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     7621 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     7681 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     7741 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     7801 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     7861 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     7921 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     7981 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     8041 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     8101 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     8161 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     8221 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     8281 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     8341 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     8401 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     8461 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     8521 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     8581 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     8641 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     8701 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     8761 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     8821 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     8881 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     8941 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     9001 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     9061 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     9121 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     9181 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     9241 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     9301 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     9361 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     9421 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     9481 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     9541 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     9601 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     9661 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     9721 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     9781 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg ccc
//
