LOCUS       sequence-349893-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing Cas9 and a sgRNA targeting UQCRC2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11590)
  AUTHORS   Spinelli JB, Rosen PC, Sprenger HG, Puszynska AM, Mann JL, Roessler 
            JM, Cangelosi AL, Henne A, Condon KJ, Zhang T, Kunchok T, Lewis CA, 
            Chandel NS, Sabatini DM
  TITLE     Fumarate is a terminal electron acceptor in the mammalian electron 
            transport chain.
  JOURNAL   Science. 2021 Dec 3;374(6572):1227-1237. doi: 
            10.1126/science.abi7495. Epub 2021 Dec 2.
  PUBMED    34855504
REFERENCE   2  (bases 1 to 11590)
  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: 
            "10.1126/science.abi7495"; journalName: "Science"; date: "2021-12-3-
            3"; volume: "374"; issue: "6572"; pages: "1227-1237"
COMMENT     Sequence Label: pLentiCRISPRv1_UQCRC2
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     404..421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    538..559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(574..579,580..597,598..604)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    612..628
                     /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)."
     primer_bind     617..639
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     636..652
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     636..652
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     671..691
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        673..691
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        719..945
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             946..1126
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1173..1298
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1791..2024
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2209..2253
                     /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"
     promoter        2431..2679
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2431..2451
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2602..2621
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2701..2776
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2824..2941
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3010..3221
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3241..3250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3250..3273
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3280..3300
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3325..7425
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3325..7425
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7426..7473
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7489..7545
                     /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             7546..8145
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7546..7565)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8042..8062
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8161..8749
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8214..8234)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8632..8643)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8821..9054
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9132..9253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9169..9188)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9223..9242
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9293..9428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9448..9467)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9449..9467)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9477..9494)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9477..9493)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9486..9508)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9635..10090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9722..9741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9932..9953
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10116..10220
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10221..10289,10290..11081)
                     /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     complement(10439..10458)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(11252..11590,1..250)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
       61 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      301 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      361 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      421 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      481 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      541 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      601 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
      661 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
      721 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
      781 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
      841 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
      901 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
      961 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     1021 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     1081 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     1141 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     1201 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1261 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1321 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1381 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     1441 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1501 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1561 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1621 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     1681 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     1741 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     1801 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     1861 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     1921 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     1981 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2041 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2101 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2161 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2221 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2281 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2341 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2401 acccacctcc caaccccgag gggacccaga gagggcctat ttcccatgat tccttcatat
     2461 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2521 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2581 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2641 ttcttggctt tatatatctt gtggaaagga cgaaacaccg taaagcaggc agtagatatg
     2701 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     2761 ggcaccgagt cggtgctttt ttgaattcta gatcttgaga caaatggcag tattcatcca
     2821 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     2881 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     2941 tcgggtttat tacagggaca gcagagatcc actttggcgc cggctcgagt ggctccggtg
     3001 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     3061 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     3121 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     3181 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtgtcgtga cgcgggatcc
     3241 gccaccatgg attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc
     3301 ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc
     3361 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     3421 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     3481 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     3541 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     3601 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     3661 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     3721 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     3781 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     3841 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     3901 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     3961 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     4021 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     4081 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     4141 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     4201 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     4261 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     4321 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     4381 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     4441 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     4501 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     4561 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     4621 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     4681 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     4741 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     4801 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     4861 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     4921 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     4981 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     5041 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     5101 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     5161 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     5221 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     5281 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     5341 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     5401 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     5461 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     5521 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     5581 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     5641 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     5701 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     5761 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     5821 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     5881 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     5941 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     6001 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     6061 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     6121 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     6181 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     6241 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     6301 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     6361 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     6421 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     6481 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     6541 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     6601 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     6661 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     6721 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     6781 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     6841 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     6901 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     6961 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     7021 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     7081 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     7141 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     7201 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     7261 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     7321 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     7381 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg tcctgctgct
     7441 actaagaaag ctggtcaagc taagaaaaag aaagctagcg gcagcggcgc caccaacttc
     7501 agcctgctga agcaggccgg cgacgtggag gagaaccccg gccccatgac cgagtacaag
     7561 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     7621 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     7681 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     7741 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     7801 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     7861 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     7921 gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     7981 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     8041 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     8101 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg ttaagtcgac
     8161 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     8221 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     8281 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     8341 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     8401 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     8461 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     8521 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     8581 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     8641 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     8701 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     8761 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     8821 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     8881 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     8941 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     9001 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtacgt
     9061 atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     9121 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     9181 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     9241 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     9301 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     9361 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     9421 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     9481 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     9541 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     9601 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     9661 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     9721 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     9781 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     9841 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     9901 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     9961 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
    10021 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
    10081 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
    10141 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
    10201 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
    10261 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
    10321 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
    10381 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
    10441 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
    10501 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
    10561 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
    10621 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
    10681 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
    10741 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
    10801 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
    10861 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
    10921 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
    10981 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
    11041 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
    11101 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
    11161 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
    11221 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
    11281 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
    11341 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
    11401 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
    11461 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
    11521 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
    11581 cgtgcacaca
//
