LOCUS       sequence-446067-       10421 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Entry vector to clone sgRNA(s) into the CROPseq-multi-v2 construct 
            with Zeocin resistance; v2 is compatible with T7 in vitro 
            transcription detection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10421)
  AUTHORS   Walton RT, Qin Y, Blainey PC
  TITLE     CROPseq-multi: a versatile solution for multiplexed perturbation and
            decoding in pooled CRISPR screens
  JOURNAL   bioRxiv 2024.03.17.585235
REFERENCE   2  (bases 1 to 10421)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.03.17.585235"
COMMENT     Sequence Label: CROPseq-multi-v2-Zeo-BsmBI_entry (pRW1500)
FEATURES             Location/Qualifiers
     source          1..10421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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    1489..1606
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1745..2923
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1976..2914
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2871..2891
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2930..2939
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2936..3310
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     misc_feature    3326..3914
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3379..3399)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3797..3808)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4049..4067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6095..6343)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(6153..6172)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6323..6343)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     LTR             6390..6570
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    6648..6769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6685..6704)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6739..6758
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6809..6944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6965..6983)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6993..7010)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6993..7009)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7002..7024)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7151..7606
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7238..7257)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7448..7469
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7632..7736
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7737..7805,7806..8597)
                     /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(7955..7974)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8768..9356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9257..9276
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9510..9527
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9644..9665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9680..9685,9686..9703,9704..9710)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9718..9734
                     /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     9723..9745
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9742..9758
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9742..9758
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9777..9797
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9779..9797
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        9825..10204
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10206..10404
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10359..10379
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa
     1501 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     1561 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     1621 ggacagcaga gatccacttt ggcgccggct cgagggggcc cgggtgcaaa gatggataaa
     1681 gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg
     1741 aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     1801 ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga
     1861 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     1921 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     1981 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     2041 aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg
     2101 gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc
     2161 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
     2221 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
     2281 ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg
     2341 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
     2401 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
     2461 gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc
     2521 accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg
     2581 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt
     2641 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
     2701 cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
     2761 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
     2821 gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca
     2881 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccatggc
     2941 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     3001 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     3061 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     3121 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     3181 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     3241 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     3301 gcaggactga acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     3361 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3421 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3481 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3541 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3601 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3661 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3721 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3781 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3841 ggctctgcgg cctcttccgc gtgttcgcct tcgccctcag acgagtcgga tctccctttg
     3901 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     3961 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagagaagat
     4021 ttcaccatta tcgtttcaga cccattttta atacgactca ctatagggaa aaaaagcacc
     4081 gactcggtgc cactttttca agttgataac ggactagcct tatttcaact tgctatgctg
     4141 ttagagacgt acaaaaaaga gcaagaagct aaaaaagatt taaaaattat ttttagcgca
     4201 gttaatggaa caggaactaa atttacccca aaaatattac gtgaatcagg atataacgtt
     4261 attgaggttg aagagcatgc atttgaagat gaaacattta aaaatgttgt aaatccaaat
     4321 ccagaatttg atcctgcatg aaaaataccg cttgaatatg gtattaaaca tgatgcagat
     4381 attattatta tgaatgaccc agatgctgac agatttggaa tggcaataaa acatgatggt
     4441 cattttgtaa gattagatgg aaatcaaaca ggaccaattt taattgattg aaaattatca
     4501 aatctaaaac gcttaaatag cattccaaaa aatccggctc tatattcaag ttttgtaaca
     4561 agtgatttgg gtgatagaat cgctcatgaa aaatatggag ttaatattgt aaaaacttta
     4621 actggattta aatgaatggg tagagaaatt gctaaagaag aagataacgg attaaatttt
     4681 gtttttgctt atgaagaaag ttatggatat gtaattgatg actcagctag agataaagat
     4741 ggaatacaag cttctatatt aatagcagag gctgcttgat tttataaaaa acaaaataaa
     4801 acattagtag actatttaga agatttattt aaagaaatgg gtgcatatta cactttcact
     4861 ttaaacttga attttaaacc agaagaaaag aaattaaaaa ttgaaccatt aatgaaatca
     4921 ttgagagcaa cacccttaac tcaaattgct ggacttaaag ttgttaatgt tgaagactac
     4981 atcgatggaa tgtataatat gccaggacaa gacttactaa aattttattt agaagataag
     5041 tcatgatttg ctgttcgccc aagtggaact gaacctaaac taaaaattta ttttataggt
     5101 gttggtgaat ctgttcaaaa cgctaaagtt aaagtagacg aaattattaa agaattaaaa
     5161 ttaaaaatga atatatagga gaaaaaatga aactaaacaa atatatagat cacacattat
     5221 taaaacaaga tgctacgaaa gctgaaatta aacaattatg tgatgaagca attgaatttg
     5281 attttgcaac agtttgtgtt aattcatatt gaacaagcta ttgtaaagaa ttattaaaag
     5341 gcacaaatgt aggaataaca aatgttgtag gttttcctct aggtgcatgc acaacagcta
     5401 caaaagcatt cgaagtttct gaagcaatta aagatggtgc aacagaaatt gatatggtat
     5461 taaatattgg tgcattaaaa gacaaaaatt atgaattagt tttagaagac atgaaagctg
     5521 taaaaaaagc agctggatca catgttgtta aatgtattat ggaaaattgt ttattaacaa
     5581 aagaagaaat catgaaagct tgtgaaatag ctgttgaagc tggattagaa tttgttaaaa
     5641 catcaacagg attttcaaaa tcaggtgcaa catttgaaga tgttaaacta atgaagtcag
     5701 ttgttaaaga caatgcttta gttaaagcag ctggtggagt tagaacattt gaagatgctc
     5761 aaaaaatgat tgaagcagga gctgaccgct taggaacaag tggtggagta gctattatta
     5821 aaggtgaaga aaacaacgcg agttactaaa actagcgttt ttttattttg ctcattttta
     5881 ttaaaagttt gcaaaaagga acataaaaat tctaattatt gatactaaag ttattaaaaa
     5941 gaagattttg gttgatttta taaaggtcat agaatataat attttagcat gtgtattttg
     6001 tgtgctcatt tacaaccgtc tctgcattgg ccgggaatcg aacccgggcc tcccgcgtgg
     6061 caggcgagaa ttctaccact gaaccaccaa tgccggtgtt tcgtcctttc cacaagatat
     6121 ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag tccattttaa
     6181 aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt cacgtatttt
     6241 gtactaatat ctttgtgttt acagtcaaat taattctaat tatctctcta acagccttgt
     6301 atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctcaagcttg gcgtaactag
     6361 atcttgagac actgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6421 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6481 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6541 ccttttagtc agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta
     6601 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6661 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6721 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6781 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6841 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6901 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
     6961 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     7021 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     7081 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     7141 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     7201 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     7261 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     7321 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     7381 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7441 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     7501 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     7561 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     7621 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     7681 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     7741 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     7801 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     7861 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     7921 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     7981 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     8041 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     8101 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     8161 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     8221 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     8281 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     8341 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     8401 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     8461 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     8521 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     8581 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     8641 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     8701 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     8761 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     8821 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     8881 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     8941 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     9001 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     9061 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     9121 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     9181 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     9241 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     9301 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     9361 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     9421 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     9481 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     9541 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     9601 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     9661 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     9721 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     9781 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaagacatt gattattgac
     9841 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     9901 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     9961 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
    10021 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
    10081 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
    10141 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
    10201 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
    10261 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
    10321 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
    10381 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac t
//
