LOCUS       sequence-297957-       13102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing high-specificity eSpCas9(1.1) and a 
            control non-targeting sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13102)
  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 13102)
  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-sgNT
FEATURES             Location/Qualifiers
     source          1..13102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1990..2238
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1990..2010
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2161..2180
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2260..2335
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2397..2608
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2623..2632
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2632..2697
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2704..2724
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2749..5286,5287..5289,5290..5751,5752..5754,5755..5922,
                     5923..5925,5926..6849)
                     /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             6850..6897
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6907..6963
                     /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             6964..7560
                     /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     7457..7477
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7576..8164
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7629..7649)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8047..8058)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8236..8469
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8495..8512)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8501..8725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8771..9199
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8858..8877)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9068..9089
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9208..9228)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9213..9542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9393..9528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9455..9474
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9590..9637
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9656..10030
                     /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    10160..10281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10197..10216)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10251..10270
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10330..10346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10330..10346)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10343..10365)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10354..10370
                     /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(10378..10384,10385..10402,10403..10408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10423..10444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10561..10578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10732..11320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10812..10831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11491..12282,12283..12351))
                     /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     12114..12133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12352..12456)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12531..12550)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12722..13101
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     2041 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
     2101 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     2161 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     2221 tggaaaggac gaaacaccga cggaggctaa gcgttgcaag ttttagagct agaaatagca
     2281 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     2341 tgaattcgct agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
     2401 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg
     2461 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     2521 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     2581 tcgcaacggg tttgccgcca gaacacagga ccggttctag atgccaccat ggactataag
     2641 gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg
     2701 gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac
     2761 agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
     2821 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
     2881 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
     2941 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
     3001 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
     3061 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
     3121 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
     3181 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
     3241 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     3301 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     3361 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     3421 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     3481 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     3541 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     3601 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     3661 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     3721 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     3781 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     3841 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     3901 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     3961 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     4021 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     4081 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     4141 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     4201 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     4261 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     4321 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     4381 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     4441 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     4501 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     4561 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     4621 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     4681 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     4741 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     4801 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     4861 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     4921 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     4981 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     5041 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     5101 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     5161 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     5221 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
     5281 tttctggccg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     5341 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     5401 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     5461 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     5521 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     5581 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     5641 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     5701 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc tgcgctggaa
     5761 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     5821 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     5881 ttcaagaccg agattaccct ggccaacggc gagatccgga aggcgcctct gatcgagaca
     5941 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     6001 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     6061 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     6121 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     6181 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     6241 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     6301 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     6361 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     6421 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     6481 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     6541 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     6601 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     6661 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     6721 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     6781 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     6841 ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggga
     6901 tccggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga
     6961 ccgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     7021 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     7081 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     7141 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     7201 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     7261 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     7321 cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc accagggcaa gggtctgggc
     7381 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     7441 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     7501 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     7561 acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     7621 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     7681 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     7741 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     7801 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     7861 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     7921 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     7981 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     8041 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     8101 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     8161 ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     8221 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     8281 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     8341 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     8401 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     8461 tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     8521 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     8581 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     8641 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     8701 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     8761 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     8821 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     8881 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     8941 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     9001 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     9061 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     9121 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     9181 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     9241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     9301 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     9361 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     9421 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     9481 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     9541 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     9601 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     9661 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     9721 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     9781 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     9841 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     9901 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     9961 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
    10021 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
    10081 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
    10141 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
    10201 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    10261 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
    10321 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
    10381 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
    10441 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
    10501 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
    10561 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
    10621 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
    10681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
    10741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
    10801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
    10861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
    10921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
    10981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
    11041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
    11101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
    11161 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
    11221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
    11281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
    11341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
    11401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
    11461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
    11521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
    11581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
    11641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
    11701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
    11761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
    11821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
    11881 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
    11941 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
    12001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
    12061 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
    12121 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
    12181 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    12241 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    12301 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    12361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    12421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    12481 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
    12541 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
    12601 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
    12661 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
    12721 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
    12781 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
    12841 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
    12901 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
    12961 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
    13021 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
    13081 ttagtcatcg ctattaccat gg
//
