LOCUS       sequence-361985-       14824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vectors with the human insulin promoter driving 
            expression of dCas9, in addition to a U6-driven sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14824)
  AUTHORS   Beucher A, Miguel-Escalada I, Balboa D, De Vas MG, Maestro MA, 
            Garcia-Hurtado J, Bernal A, Gonzalez-Franco R, Vargiu P, Heyn H, 
            Ravassard P, Ortega S, Ferrer J
  TITLE     The HASTER lncRNA promoter is a cis-acting transcriptional 
            stabilizer of HNF1A.
  JOURNAL   Nat Cell Biol. 2022 Oct 6. pii: 10.1038/s41556-022-00996-8. doi: 
            10.1038/s41556-022-00996-8.
  PUBMED    36202974
REFERENCE   2  (bases 1 to 14824)
  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: "Nat Cell 
            Biol. 2022 Oct 6. pii: 10.1038/s41556-022-00996-8. doi: 
            10.1038/s41556-022-00996-8."
COMMENT     Sequence Label: pLV-hIP-dCas9-BSD
FEATURES             Location/Qualifiers
     source          1..14824
                     /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        4120..4195
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     4232..4250
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     regulatory      4597..4606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4603..4629,4630..4632,4633..7188,7189..7191,
                     7192..8706)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8761..8781
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8821..8874
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             8875..9273
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9298..9886
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9351..9371)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9769..9780)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9958..10191
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10217..10234)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10223..10447
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10493..10921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10580..10599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10790..10811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10930..10950)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10935..11264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11115..11250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11177..11196
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11312..11359
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11378..11752
                     /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    11882..12003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11919..11938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11973..11992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12052..12068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12052..12068)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12065..12087)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12076..12092
                     /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(12100..12106,12107..12124,12125..12130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12145..12166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12283..12300)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12454..13042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12534..12553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13213..14004,14005..14073))
                     /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     13836..13855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14074..14178)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14253..14272)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        14444..14823
                     /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 gaaacaccgg agacggttgt aaatgagcac acaaaataca catgctaaaa
     2281 tattatattc tatgaccttt ataaaatcaa ccaaaatctt ctttttaata actttagtat
     2341 caataattag aatttttatg ttcctttttg caaactttta ataaaaatga gcaaaataaa
     2401 aaaacgctag ttttagtaac tcgcgttgtt ttcttcacct ttaataatag ctactccacc
     2461 acttgttcct aagcggtcag ctcctgcttc aatcattttt tgagcatctt caaatgttct
     2521 aactccacca gctgctttaa ctaaagcatt gtctttaaca actgacttca ttagtttaac
     2581 atcttcaaat gttgcacctg attttgaaaa tcctgttgat gttttaacaa attctaatcc
     2641 agcttcaaca gctatttcac aagctttcat gatttcttct tttgttaata aacaattttc
     2701 cataatacat ttaacaacat gtgatccagc tgcttttttt acagctttca tgtcttctaa
     2761 aactaattca taatttttgt cttttaatgc accaatattt aataccatat caatttctgt
     2821 tgcaccatct ttaattgctt cagaaacttc gaatgctttt gtagctgttg tgcatgcacc
     2881 tagaggaaaa cctacaacat ttgttattcc tacatttgtg ccttttaata attctttaca
     2941 atagcttgtt caatatgaat taacacaaac tgttgcaaaa tcaaattcaa ttgcttcatc
     3001 acataattgt ttaatttcag ctttcgtagc atcttgtttt aataatgtgt gatctatata
     3061 tttgtttagt ttcatttttt ctcctatata ttcattttta attttaattc tttaataatt
     3121 tcgtctactt taactttagc gttttgaaca gattcaccaa cacctataaa ataaattttt
     3181 agtttaggtt cagttccact tgggcgaaca gcaaatcatg acttatcttc taaataaaat
     3241 tttagtaagt cttgtcctgg catattatac attccatcga tgtagtcttc aacattaaca
     3301 actttaagtc cagcaatttg agttaagggt gttgctctca atgatttcat taatggttca
     3361 atttttaatt tcttttcttc tggtttaaaa ttcaagttta aagtgaaagt gtaatatgca
     3421 cccatttctt taaataaatc ttctaaatag tctactaatg ttttattttg ttttttataa
     3481 aatcaagcag cctctgctat taatatagaa gcttgtattc catctttatc tctagctgag
     3541 tcatcaatta catatccata actttcttca taagcaaaaa caaaatttaa tccgttatct
     3601 tcttctttag caatttctct acccattcat ttaaatccag ttaaagtttt tacaatatta
     3661 actccatatt tttcatgagc gattctatca cccaaatcac ttgttacaaa acttgaatat
     3721 agagccggat tttttggaat gctatttaag cgttttagat ttgataattt tcaatcaatt
     3781 aaaattggtc ctgtttgatt tccatctaat cttacaaaat gaccatcatg ttttattgcc
     3841 attccaaatc tgtcagcatc tgggtcattc ataataataa tatctgcatc atgtttaata
     3901 ccatattcaa gcggtatttt tcatgcagga tcaaattctg gatttggatt tacaacattt
     3961 ttaaatgttt catcttcaaa tgcatgctct tcaacctcaa taacgttata tcctgattca
     4021 cgtaatattt ttggggtaaa tttagttcct gttccattaa ctgcgctaaa aataattttt
     4081 aaatcttttt tagcttcttg ctcttttttg tacgtctctg ttttagagct agaaatagca
     4141 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     4201 tgaattcgtt atcatgacat taacctataa aaataggcgt atcacgaggc cctgctctcc
     4261 tggtctaatg tggaaagtgg cccaggtgag ggctttgctc tcctggagac atttgccccc
     4321 agctgtgagc agggacaggt ctggccaccg ggcccctggt taagactcta atgacccgct
     4381 ggtcctgagg aagaggtgct gacgaccaag gagatcttcc cacagaccca gcaccaggga
     4441 aatggtccgg aaattgcagc ctcagccccc agccatctgc cgaccccccc accccaggcc
     4501 ctaatgggcc aggcggcagg ggttgacagg taggggagat gggctctgag actataaagc
     4561 cagcgggggc ccagcagccc tcagccctcc aggacagcca ccatggacaa gaagtacagc
     4621 atcggcctgg ccatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     4681 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     4741 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     4801 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     4861 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     4921 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     4981 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     5041 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     5101 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     5161 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     5221 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     5281 gcccagctgc ccggcgagaa gaagaatggc ctgttcggca acctgattgc cctgagcctg
     5341 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     5401 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     5461 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     5521 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     5581 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     5641 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     5701 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     5761 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     5821 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     5881 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     5941 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     6001 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     6061 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     6121 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     6181 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     6241 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     6301 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     6361 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     6421 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     6481 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     6541 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     6601 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     6661 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     6721 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     6781 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     6841 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     6901 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     6961 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     7021 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     7081 gaactggaca tcaaccggct gtccgactac gatgtggacc acatcgtgcc tcagagcttt
     7141 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaaggc ccggggcaag
     7201 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     7261 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     7321 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     7381 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     7441 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     7501 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     7561 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     7621 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     7681 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     7741 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     7801 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     7861 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     7921 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     7981 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     8041 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     8101 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     8161 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     8221 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     8281 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     8341 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     8401 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     8461 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     8521 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     8581 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     8641 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     8701 ggcgacagcg ctggaggagg tggaagcgga ggaggaggaa gcggaggagg aggtagcgga
     8761 cctaagaaaa agaggaaggt ggcggccgct ggatccggac gggctggtac aggcagtgga
     8821 gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg cccaatggcc
     8881 aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc
     8941 atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc
     9001 ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg
     9061 ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag
     9121 aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct
     9181 gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa
     9241 ttgctgccct ctggttatgt gtgggagggc taagaattcg atatcaagct tatcggtaat
     9301 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     9361 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     9421 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     9481 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     9541 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     9601 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     9661 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     9721 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     9781 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     9841 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     9901 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     9961 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
    10021 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
    10081 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
    10141 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt
    10201 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
    10261 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
    10321 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
    10381 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
    10441 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
    10501 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
    10561 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
    10621 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
    10681 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
    10741 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
    10801 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
    10861 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
    10921 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
    10981 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
    11041 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
    11101 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
    11161 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
    11221 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
    11281 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
    11341 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
    11401 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
    11461 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
    11521 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
    11581 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
    11641 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
    11701 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
    11761 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
    11821 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
    11881 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
    11941 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
    12001 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
    12061 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    12121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    12181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    12241 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    12301 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    12361 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    12421 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    12481 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    12541 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    12601 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    12661 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    12721 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    12781 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    12841 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    12901 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    12961 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    13021 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    13081 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    13141 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    13201 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    13261 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    13321 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    13381 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    13441 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    13501 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    13561 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    13621 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    13681 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    13741 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    13801 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    13861 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    13921 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    13981 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    14041 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    14101 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    14161 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
    14221 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
    14281 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
    14341 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
    14401 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
    14461 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
    14521 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
    14581 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
    14641 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
    14701 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
    14761 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
    14821 atgg
//
