LOCUS       sequence-297967-       13101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing high-specificity eSpCas9(1.1) and a 
            gRNA targeting human MIB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13101)
  AUTHORS   Bauer M, Flatt JW, Seiler D, Cardel B, Emmenlauer M, Boucke K, 
            Suomalainen M, Hemmi S, Greber UF
  TITLE     The E3 Ubiquitin Ligase Mind Bomb 1 Controls Adenovirus Genome 
            Release at the Nuclear Pore Complex.
  JOURNAL   Cell Rep. 2019 Dec 17;29(12):3785-3795.e8. doi: 
            10.1016/j.celrep.2019.11.064.
  PUBMED    31851912
REFERENCE   2  (bases 1 to 13101)
  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"; doi: 
            "10.1016/j.celrep.2019.11"; journalName: "Cell Rep"; date: 
            "2019-12-17- 17"; volume: "29"; issue: "12"; pages: "3785-3795.e8"
COMMENT     Sequence Label: Lenti-eCas9-sgMIB1
FEATURES             Location/Qualifiers
     source          1..13101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2371..2619
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2371..2391
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2542..2561
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2640..2715
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2777..2988
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3003..3012
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3012..3077
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3084..3104
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3129..5666,5667..5669,5670..6131,6132..6134,6135..6302,
                     6303..6305,6306..7229)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7230..7277
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7287..7343
                     /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             7344..7940
                     /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     7837..7857
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7956..8544
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8009..8029)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8427..8438)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8616..8849
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8875..8892)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8881..9105
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9151..9579
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9238..9257)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9448..9469
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9588..9608)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9593..9922
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9773..9908
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9835..9854
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9970..10017
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10036..10410
                     /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    10540..10661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10577..10596)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10631..10650
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10710..10726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10710..10726)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10723..10745)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10734..10750
                     /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(10758..10764,10765..10782,10783..10788))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10803..10824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10941..10958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11112..11700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11192..11211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11871..12662,12663..12731))
                     /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     12494..12513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12732..12836)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12911..12930)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat
     2401 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2461 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2521 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2581 ttcttggctt tatatatctt gtggaaagga cgaaacaccg ttggcgctcg ggtagtgcgg
     2641 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     2701 gcaccgagtc ggtgcttttt tgaattcgct agctaggtct tgaaaggagt gggaattggc
     2761 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     2821 ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     2881 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     2941 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagga ccggttctag
     3001 atgccaccat ggactataag gaccacgacg gagactacaa ggatcatgat attgattaca
     3061 aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc
     3121 cagcagccga caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg
     3181 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg
     3241 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag
     3301 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga
     3361 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc
     3421 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct
     3481 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga
     3541 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg
     3601 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca
     3661 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa
     3721 accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga
     3781 gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg
     3841 gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg
     3901 ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc
     3961 tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg
     4021 ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg
     4081 cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg
     4141 tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct
     4201 acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca
     4261 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc
     4321 tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc
     4381 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa
     4441 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa
     4501 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg
     4561 aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg
     4621 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca
     4681 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct
     4741 tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag
     4801 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg
     4861 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga
     4921 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata
     4981 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct
     5041 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct
     5101 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa
     5161 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg
     5221 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata
     5281 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
     5341 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
     5401 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
     5461 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
     5521 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
     5581 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
     5641 accatatcgt gcctcagagc tttctggccg acgactccat cgacaacaag gtgctgacca
     5701 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
     5761 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
     5821 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
     5881 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
     5941 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
     6001 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
     6061 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
     6121 aaaagtaccc tgcgctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
     6181 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
     6241 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
     6301 aggcgcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
     6361 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
     6421 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc
     6481 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg
     6541 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
     6601 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca
     6661 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc
     6721 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg
     6781 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
     6841 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
     6901 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
     6961 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
     7021 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
     7081 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca
     7141 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
     7201 ggatcgacct gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc
     7261 aggcaaaaaa gaaaaaggga tccggcgcaa caaacttctc tctgctgaaa caagccggag
     7321 atgtcgaaga gaatcctgga ccgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     7381 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     7441 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     7501 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     7561 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     7621 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     7681 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc
     7741 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     7801 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     7861 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     7921 cccgcaagcc cggtgcctga acgcgttaag tcgacaatca acctctggat tacaaaattt
     7981 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     8041 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     8101 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     8161 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     8221 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     8281 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     8341 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     8401 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     8461 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     8521 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
     8581 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     8641 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     8701 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     8761 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     8821 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
     8881 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     8941 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     9001 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     9061 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     9121 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     9181 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     9241 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     9301 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     9361 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     9421 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     9481 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     9541 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     9601 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     9661 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     9721 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     9781 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     9841 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     9901 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     9961 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
    10021 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
    10081 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
    10141 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
    10201 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
    10261 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
    10321 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
    10381 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
    10441 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
    10501 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
    10561 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
    10621 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
    10681 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
    10741 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
    10801 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
    10861 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
    10921 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
    10981 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
    11041 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
    11101 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
    11161 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
    11221 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
    11281 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
    11341 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
    11401 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
    11461 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
    11521 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
    11581 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
    11641 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
    11701 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
    11761 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
    11821 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
    11881 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
    11941 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
    12001 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
    12061 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
    12121 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
    12181 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
    12241 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    12301 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    12361 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    12421 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    12481 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    12541 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    12601 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    12661 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    12721 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    12781 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    12841 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
    12901 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
    12961 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
    13021 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
    13081 tacgggccag atatacgcgt t
//
