LOCUS       sequence-171534-       13012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus carrying Cas9/CRISPR for cut in GFP (used as control).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13012)
  AUTHORS   Huang Y, Tejero R, Lee VK, Brusco C, Hannah T, Bertucci TB, 
            Junqueira Alves C, Katsyv I, Kluge M, Foty R, Zhang B, Friedel CC, 
            Dai G, Zou H, Friedel RH
  TITLE     Plexin-B2 facilitates glioblastoma infiltration by modulating cell 
            biomechanics.
  JOURNAL   Commun Biol. 2021 Jan 29;4(1):145. doi: 10.1038/s42003-021-01667-4.
  PUBMED    33514835
REFERENCE   2  (bases 1 to 13012)
  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: "Commun 
            Biol."; date: "2021-01-29"; volume: "4(1):145. doi"; pages: " 
            10.1038/s42003-021-01667-4"
COMMENT     Sequence Label: plentiCRISPRv2-sgEGFP
FEATURES             Location/Qualifiers
     source          1..13012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..36)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(190..778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(270..289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(949..1740,1741..1809))
                     /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     1572..1591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1810..1914)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1989..2008)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2180..2559
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2561..2759
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2714..2734
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2777..2957
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3004..3129
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3622..3855
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4040..4084
                     /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    4382..4499
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4550..4798
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4550..4570
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4721..4740
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4819..4894
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4956..5167
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5186..5195
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5192..9295
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5192..9295
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9296..9343
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9344..9367
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9377..9433
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             9434..10030
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9927..9947
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10046..10634
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10099..10119)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10517..10528)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10706..10939
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10965..10982)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10971..11195
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11241..11669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11328..11347)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11538..11559
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11678..11698)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11683..12012
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11863..11998
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11925..11944
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12060..12107
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12126..12500
                     /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    12630..12751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12667..12686)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12721..12740
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12800..12816)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12800..12816)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12813..12835)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12824..12840
                     /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(12848..12854,12855..12872,12873..12878))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12893..12914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
       61 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1921 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     1981 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     2041 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     2101 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     2161 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     2221 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     2281 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     2341 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2401 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     2461 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     2521 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2581 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     2641 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     2701 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc
     2761 gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     2821 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     2881 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     2941 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
     3001 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     3061 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     3121 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     3181 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     3241 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     3301 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     3361 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     3421 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     3481 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     3541 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     3601 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     3661 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     3721 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     3781 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     3841 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     3901 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     3961 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     4021 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     4081 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     4141 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     4201 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     4261 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     4321 agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca
     4381 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     4441 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     4501 gggtttatta cagggacagc agagatccag tttggttaat taaggtaccg agggcctatt
     4561 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattagaat
     4621 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     4681 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     4741 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg
     4801 gcgaggagct gttcaccggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
     4861 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gaattcgcta gctaggtctt
     4921 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     4981 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
     5041 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     5101 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     5161 aacacaggac cggttctaga gcgctgccac catggacaag aagtacagca tcggcctgga
     5221 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     5281 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     5341 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     5401 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     5461 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     5521 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     5581 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     5641 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     5701 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     5761 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     5821 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     5881 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     5941 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     6001 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     6061 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     6121 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     6181 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     6241 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     6301 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     6361 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     6421 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     6481 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     6541 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     6601 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     6661 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     6721 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     6781 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     6841 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     6901 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     6961 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     7021 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     7081 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     7141 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     7201 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     7261 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     7321 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     7381 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     7441 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     7501 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     7561 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     7621 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     7681 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     7741 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     7801 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     7861 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     7921 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     7981 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     8041 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     8101 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     8161 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     8221 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     8281 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     8341 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     8401 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     8461 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     8521 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     8581 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     8641 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     8701 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     8761 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     8821 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     8881 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     8941 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     9001 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     9061 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     9121 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     9181 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     9241 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg
     9301 acctgccgcc acaaagaagg ctggacaggc taagaagaag aaagattaca aagacgatga
     9361 cgataaggga tccggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga
     9421 gaatcctgga ccgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     9481 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     9541 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     9601 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     9661 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     9721 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     9781 gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc accagggcaa
     9841 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     9901 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     9961 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
    10021 cggtgcctga acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
    10081 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
    10141 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
    10201 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
    10261 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
    10321 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
    10381 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
    10441 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
    10501 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
    10561 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
    10621 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
    10681 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
    10741 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
    10801 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
    10861 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
    10921 gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
    10981 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
    11041 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
    11101 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
    11161 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
    11221 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
    11281 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
    11341 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
    11401 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
    11461 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
    11521 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
    11581 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
    11641 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
    11701 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
    11761 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
    11821 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
    11881 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
    11941 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
    12001 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
    12061 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
    12121 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
    12181 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
    12241 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
    12301 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
    12361 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
    12421 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
    12481 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
    12541 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
    12601 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
    12661 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
    12721 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
    12781 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
    12841 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
    12901 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
    12961 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt at
//
