LOCUS       sequence-424959-       10799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector to clone sgRNA(s) into the CROPseq-multi construct with
            nuclear-localized EBFP2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10799)
  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 10799)
  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 
            2024.03.17.585235"
COMMENT     Sequence Label: CROPseq-multi-EBFP2-NLS (pRW1447)
FEATURES             Location/Qualifiers
     source          1..10799
                     /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             join(2936..2938,2939..2941,2942..3652)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHLKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(2981..3002)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3665..3685
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3704..4292
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3757..3777)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4175..4186)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(6473..6721)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(6531..6550)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6701..6721)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     LTR             6768..6948
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    7026..7147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7063..7082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7117..7136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7187..7322
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7342..7361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7343..7361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7371..7388)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7371..7387)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7380..7402)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7529..7984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7616..7635)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7826..7847
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8010..8114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8115..8183,8184..8975)
                     /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(8333..8352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9146..9734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9635..9654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9888..9905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10022..10043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10058..10063,10064..10081,10082..10088)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10096..10112
                     /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     10101..10123
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10120..10136
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10120..10136
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10155..10175
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10157..10175
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        10203..10582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10584..10782
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10737..10757
                     /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 ccaccatggt
     2941 cagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     3001 cgtcaacggc cacaagttca gcgtcagggg cgagggcgag ggcgacgcca ccaacggcaa
     3061 gctcaccctc aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     3121 caccaccctc agccacggcg tgcagtgctt cgcccgctac cccgaccacc tcaagcagca
     3181 cgacttcttc aagtccgcaa tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3241 ggacgacggc acctacaaga cccgcgccga ggtcaagttc gagggcgaca ccctggtcaa
     3301 ccgcatcgag ctcaagggcg tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3361 ggagtacaac ttcaacagcc acaacatcta tattatggcc gtcaagcaga agaacggcat
     3421 caaggtcaac ttcaagatcc gccacaacgt ggaggacggc agcgtgcagc tcgccgacca
     3481 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca gccactacct
     3541 cagcacccag tccgtgctca gcaaagaccc caacgagaag cgcgatcata tggtcctgct
     3601 ggagttccgc accgccgccg ggatcactct cggtatggac gagctctaca agtctggtgg
     3661 ttctcccaag aagaagagga aagtctaaac gcgttaagtc gacaatcaac ctctggatta
     3721 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3781 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3841 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3901 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3961 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4021 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4081 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     4141 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4201 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt gttcgccttc gccctcagac
     4261 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
     4321 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     4381 tcccaacgaa gagaagattt caccattatc gtttcagacc cacctcccaa ccccgagggg
     4441 acccagaaaa aaagcaccga ctcggtgcca ctttttcaag ttgataacgg actagcctta
     4501 tttcaacttg ctatgctgtt agagacgtac aaaaaagagc aagaagctaa aaaagattta
     4561 aaaattattt ttagcgcagt taatggaaca ggaactaaat ttaccccaaa aatattacgt
     4621 gaatcaggat ataacgttat tgaggttgaa gagcatgcat ttgaagatga aacatttaaa
     4681 aatgttgtaa atccaaatcc agaatttgat cctgcatgaa aaataccgct tgaatatggt
     4741 attaaacatg atgcagatat tattattatg aatgacccag atgctgacag atttggaatg
     4801 gcaataaaac atgatggtca ttttgtaaga ttagatggaa atcaaacagg accaatttta
     4861 attgattgaa aattatcaaa tctaaaacgc ttaaatagca ttccaaaaaa tccggctcta
     4921 tattcaagtt ttgtaacaag tgatttgggt gatagaatcg ctcatgaaaa atatggagtt
     4981 aatattgtaa aaactttaac tggatttaaa tgaatgggta gagaaattgc taaagaagaa
     5041 gataacggat taaattttgt ttttgcttat gaagaaagtt atggatatgt aattgatgac
     5101 tcagctagag ataaagatgg aatacaagct tctatattaa tagcagaggc tgcttgattt
     5161 tataaaaaac aaaataaaac attagtagac tatttagaag atttatttaa agaaatgggt
     5221 gcatattata ctttcacttt aaacttgaat tttaaaccag aagaaaagaa attaaaaatt
     5281 gaaccattaa tgaaatcatt gagagcaaca cccttaactc aaattgctgg acttaaagtt
     5341 gttaatgttg aagactacat cgatggaatg tataatatgc caggacaaga cttactaaaa
     5401 ttttatttag aagataagtc atgatttgct gttcgcccaa gtggaactga acctaaacta
     5461 aaaatttatt ttataggtgt tggtgaatct gttcaaaacg ctaaagttaa agtagacgaa
     5521 attattaaag aattaaaatt aaaaatgaat atataggaga aaaaatgaaa ctaaacaaat
     5581 atatagatca cacattatta aaacaagatg ctacgaaagc tgaaattaaa caattatgtg
     5641 atgaagcaat tgaatttgat tttgcaacag tttgtgttaa ttcatattga acaagctatt
     5701 gtaaagaatt attaaaaggc acaaatgtag gaataacaaa tgttgtaggt tttcctctag
     5761 gtgcatgcac aacagctaca aaagcattcg aagtttctga agcaattaaa gatggtgcaa
     5821 cagaaattga tatggtatta aatattggtg cattaaaaga caaaaattat gaattagttt
     5881 tagaagacat gaaagctgta aaaaaagcag ctggatcaca tgttgttaaa tgtattatgg
     5941 aaaattgttt attaacaaaa gaagaaatca tgaaagcttg tgaaatagct gttgaagctg
     6001 gattagaatt tgttaaaaca tcaacaggat tttcaaaatc aggtgcaaca tttgaagatg
     6061 ttaaactaat gaagtcagtt gttaaagaca atgctttagt taaagcagct ggtggagtta
     6121 gaacatttga agatgctcaa aaaatgattg aagcaggagc tgaccgctta ggaacaagtg
     6181 gtggagtagc tattattaaa ggtgaagaaa acaacgcgag ttactaaaac tagcgttttt
     6241 ttattttgct catttttatt aaaagtttgc aaaaaggaac ataaaaattc taattattga
     6301 tactaaagtt attaaaaaga agattttggt tgattttata aaggtcatag aatataatat
     6361 tttagcatgt gtattttgtg tgctcattta caaccgtctc tgcattggcc gggaatcgaa
     6421 cccgggcctc ccgcgtggca ggcgagaatt ctaccactga accaccaatg ccggtgtttc
     6481 gtcctttcca caagatatat aaagccaaga aatcgaaata ctttcaagtt acggtaagca
     6541 tatgatagtc cattttaaaa cataatttta aaactgcaaa ctacccaaga aattattact
     6601 ttctacgtca cgtattttgt actaatatct ttgtgtttac agtcaaatta attctaatta
     6661 tctctctaac agccttgtat cgtatatgca aatatgaagg aatcatggga aataggccct
     6721 caagcttggc gtaactagat cttgagacac tgctttttgc ttgtactggg tctctctggt
     6781 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6841 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6901 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta
     6961 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     7021 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     7081 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     7141 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     7201 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     7261 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     7321 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     7381 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     7441 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     7501 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     7561 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     7621 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     7681 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     7741 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     7801 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     7861 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     7921 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     7981 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     8041 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     8101 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     8161 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     8221 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     8281 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     8341 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     8401 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     8461 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     8521 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     8581 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     8641 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     8701 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     8761 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     8821 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     8881 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     8941 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     9001 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     9061 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     9121 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     9181 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     9241 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     9301 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     9361 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     9421 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     9481 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     9541 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     9601 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     9661 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     9721 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     9781 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     9841 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     9901 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     9961 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
    10021 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
    10081 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
    10141 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt
    10201 aagacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
    10261 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
    10321 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
    10381 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
    10441 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
    10501 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
    10561 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
    10621 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
    10681 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
    10741 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtact
//
