LOCUS       sequence-388050-       14231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-KO. WT-SpCas9 and sgRNA targeting HPRT1. Editing-competent 
            cells can be selected with 6-TG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14231)
  AUTHORS   Schmierer B, Botla SK, Zhang J, Turunen M, Kivioja T, Taipale J
  TITLE     CRISPR/Cas9 screening using unique molecular identifiers.
  JOURNAL   Mol Syst Biol. 2017 Oct 9;13(10):945. doi: 10.15252/msb.20177834.
  PUBMED    28993443
REFERENCE   2  (bases 1 to 14231)
  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: "Mol Syst 
            Biol."; date: "2017-10-9"; volume: "13(10):945. doi"; pages: " 
            10.15252/msb.20177834"
COMMENT     Sequence Label: lenti-Cas9-sgHPRT1
FEATURES             Location/Qualifiers
     source          1..14231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..43)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    32..256
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      302..730
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(389..408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     599..620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(739..759)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        744..1073
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      924..1059
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     986..1005
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1121..1168
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1187..1561
                     /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"
     polyA_signal    1691..1812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1728..1747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1782..1801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1861..1877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1861..1877)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1874..1896)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1885..1901
                     /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(1909..1915,1916..1933,1934..1939))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1954..1975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2092..2109)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2263..2851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2343..2362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3022..3813,3814..3882))
                     /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     3645..3664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3883..3987)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4062..4081)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4253..4632
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4634..4832
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4787..4807
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4850..5030
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5077..5202
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5695..5928
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6113..6157
                     /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    6455..6572
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6702..7880
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6933..7871
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7828..7848
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7896..7943
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7944..12044
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7944..12044
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             12051..12071
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12096..12149
                     /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             12150..12548
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     12731..12750
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        12829..12904
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    12938..13526
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12991..13011)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13409..13420)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(13529..13545)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13530..13546)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             14051..14231
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
       61 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
      121 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
      181 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
      241 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
      301 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      361 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      421 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      481 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      541 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      601 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      661 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      721 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      781 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      841 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      901 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
      961 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     1021 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     1081 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     1141 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
     1201 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
     1261 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
     1321 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
     1381 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
     1441 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
     1501 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
     1561 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     1621 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     1681 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1741 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1801 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     1861 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1921 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1981 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2041 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2101 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2161 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2221 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2281 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2341 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2401 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2461 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2521 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2581 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2641 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2701 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2761 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2821 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2881 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2941 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3001 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3061 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3121 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3181 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3241 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3301 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3361 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3421 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3481 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3541 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3601 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3661 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3721 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3781 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3841 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3901 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3961 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     4021 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     4081 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     4141 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     4201 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     4261 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     4321 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     4381 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     4441 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     4501 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     4561 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     4621 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     4681 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     4741 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     4801 aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg ggtctctctg
     4861 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     4921 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     4981 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
     5041 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
     5101 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
     5161 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     5221 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     5281 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     5341 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     5401 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     5461 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     5521 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
     5581 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     5641 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     5701 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     5761 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     5821 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     5881 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     5941 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     6001 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     6061 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     6121 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     6181 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     6241 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     6301 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     6361 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc
     6421 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     6481 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     6541 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     6601 cagtttggtt aattagctag ctgcaaagat ggataaagtt ttaaacagag aggaatcttt
     6661 gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt
     6721 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     6781 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     6841 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     6901 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     6961 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
     7021 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
     7081 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
     7141 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     7201 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     7261 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     7321 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     7381 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     7441 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     7501 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
     7561 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     7621 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
     7681 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     7741 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     7801 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     7861 ttccatttca ggtgtcgtga cgtacggcca ccatgaaaag gccggcggcc acgaaaaagg
     7921 ccggccaggc aaaaaagaaa aaggacaaga agtacagcat cggcctggac atcggcacca
     7981 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg
     8041 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg
     8101 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca
     8161 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg
     8221 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg
     8281 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
     8341 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga
     8401 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc
     8461 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     8521 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     8581 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga
     8641 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     8701 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     8761 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     8821 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     8881 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     8941 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     9001 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca
     9061 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     9121 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     9181 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
     9241 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
     9301 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
     9361 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc
     9421 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
     9481 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa
     9541 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca
     9601 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
     9661 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     9721 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     9781 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     9841 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     9901 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     9961 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
    10021 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
    10081 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
    10141 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
    10201 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
    10261 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
    10321 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
    10381 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
    10441 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca
    10501 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
    10561 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc
    10621 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
    10681 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
    10741 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga
    10801 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
    10861 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
    10921 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
    10981 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
    11041 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
    11101 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt
    11161 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata
    11221 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
    11281 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
    11341 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
    11401 ccaagaaact gaagagtgtg aaagagctac tggggatcac catcatggaa agaagcagct
    11461 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
    11521 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
    11581 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
    11641 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
    11701 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca
    11761 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct
    11821 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta
    11881 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
    11941 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
    12001 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacagcgga cctaagaaaa
    12061 agaggaaggt ggcggccgct ggatccggca gtggagaggg cagaggaagt ctgctaacat
    12121 gcggtgacgt cgaggagaat cctggcccaa tggccaagcc tttgtctcaa gaagaatcca
    12181 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
    12241 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
    12301 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
    12361 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
    12421 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
    12481 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
    12541 agggctaaga tatctagatt atcggtaatt tcccatgatt ccttcatatt tgcatatacg
    12601 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
    12661 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
    12721 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
    12781 atatatcttg tggaaaggac gaaacaccga tgtgatgaag gagatggggt tttagagcta
    12841 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
    12901 gtgctttttt ctagaattcg atatcaagct tatcggtaat caacctctgg attacaaaat
    12961 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
    13021 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
    13081 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
    13141 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
    13201 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
    13261 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
    13321 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
    13381 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
    13441 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
    13501 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca
    13561 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
    13621 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
    13681 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
    13741 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
    13801 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
    13861 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
    13921 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
    13981 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
    14041 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
    14101 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
    14161 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
    14221 aatctctagc a
//
