LOCUS       sequence-398297-       13332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a CRISPRi guide targeting BIRC6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13332)
  AUTHORS   Cervia LD, Shibue T, Borah AA, Gaeta B, He L, Leung L, Li N, Moyer 
            SM, Shim BH, Dumont N, Gonzalez A, Bick NR, Kazachkova M, Dempster 
            JM, Krill-Burger JM, Piccioni F, Udeshi ND, Olive ME, Carr SA, Root 
            DE, McFarland JM, Vazquez F, Hahn WC
  TITLE     A Ubiquitination Cascade Regulating the Integrated Stress Response 
            and Survival in Carcinomas.
  JOURNAL   Cancer Discov. 2023 Mar 1;13(3):766-795. doi: 
            10.1158/2159-8290.CD-22-1230.
  PUBMED    36576405
REFERENCE   2  (bases 1 to 13332)
  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: "10.1158/2159-8290";
            journalName: "Cancer Discov"; date: "2023-03-1- 1"; volume: "13"; 
            issue: "3"; pages: "766-795"
COMMENT     Sequence Label: pXPR_023d-sgCiBIRC6-5
FEATURES             Location/Qualifiers
     source          1..13332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..43)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        215..594
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        596..794
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     749..769
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             812..992
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1039..1164
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1657..1890
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2075..2119
                     /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    2417..2534
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2585..2833
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2585..2605
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2756..2775
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2855..2930
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2992..3203
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    3229..3253
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3283..3468
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             join(3490..3516,3517..3519,3520..6006,6007..6009,
                     6010..7593)
                     /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 H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3490..7593
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7597..7623
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7603..7629
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             7642..7662
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7669..7689
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7732..7788
                     /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             7789..8385
                     /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     8282..8302
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8401..8989
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8454..8474)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8872..8883)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9061..9294
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9320..9337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9326..9550
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9596..10024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9683..9702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9893..9914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10033..10053)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10038..10367
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10218..10353
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10280..10299
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10415..10462
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10481..10855
                     /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    10985..11106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11022..11041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11076..11095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11155..11171)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11155..11171)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11168..11190)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11179..11195
                     /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(11203..11209,11210..11227,11228..11233))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11248..11269
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11386..11403)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11557..12145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11637..11656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12316..13107,13108..13176))
                     /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     12939..12958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13177..13281)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
       61 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
      121 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
      181 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
      241 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      301 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      361 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      421 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      481 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      541 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      601 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      661 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      721 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      781 gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc agatctgagc
      841 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
      901 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
      961 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
     1021 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
     1081 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
     1141 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
     1201 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
     1261 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
     1321 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     1381 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
     1441 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
     1501 gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     1561 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     1621 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     1681 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     1741 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1801 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1861 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1921 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1981 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     2041 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     2101 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     2161 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     2221 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     2281 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     2341 cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg
     2401 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     2461 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     2521 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattaagg
     2581 taccgagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     2641 agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     2701 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     2761 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
     2821 aggacgaaac accggggccc cggctgctcc tcgggtttta gagctagaaa tagcaagtta
     2881 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgaat
     2941 tcgctagcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc
     3001 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga tccggtgcct
     3061 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     3121 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     3181 acgggtttgc cgccagaaca caggaccggt tctagcatcg attgatcaac aagtttgtac
     3241 aaaaaagttg gcatggatgc taagtcacta actgcctggt cccggacact ggtgaccttc
     3301 aaggatgtat ttgtggactt caccagggag gagtggaagc tgctggacac tgctcagcag
     3361 atcgtgtaca gaaatgtgat gctggagaac tataagaacc tggtttcctt gggttatcag
     3421 cttactaagc cagatgtgat cctccggttg gagaagggag aagagcccgg cggttccggc
     3481 ggagggtcga tggacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc
     3541 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     3601 accgaccggc acagcatcaa gaagaacctg atcggcgccc tgctgttcga cagcggagaa
     3661 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     3721 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     3781 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     3841 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     3901 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     3961 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     4021 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     4081 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     4141 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     4201 ttcggcaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     4261 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     4321 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     4381 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     4441 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     4501 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     4561 ggctacgccg gctacatcga tggcggagcc agccaggaag agttctacaa gttcatcaag
     4621 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     4681 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     4741 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     4801 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     4861 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     4921 ttcgaggaag tggtggacaa gggcgccagc gcccagagct tcatcgagcg gatgaccaac
     4981 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     5041 ttcaccgtgt acaacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     5101 gccttcctga gcggcgagca gaaaaaagcc atcgtggacc tgctgttcaa gaccaaccgg
     5161 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     5221 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     5281 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     5341 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     5401 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     5461 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     5521 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     5581 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     5641 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     5701 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     5761 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     5821 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     5881 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     5941 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     6001 gtggacgcta tcgtgcctca gagctttctg aaggacgact ccatcgataa caaagtgctg
     6061 actcggagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     6121 aagatgaaga actactggcg ccagctgctg aatgccaagc tgattaccca gaggaagttc
     6181 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     6241 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     6301 cggatgaaca ctaagtacga cgagaacgac aaactgatcc gggaagtgaa agtgatcacc
     6361 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     6421 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     6481 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     6541 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     6601 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     6661 cggaagcggc ctctgatcga gacaaacggc gaaacaggcg agatcgtgtg ggataagggc
     6721 cgggactttg ccaccgtgcg gaaagtgctg tctatgcccc aagtgaatat cgtgaaaaag
     6781 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgac
     6841 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     6901 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     6961 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     7021 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     7081 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     7141 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     7201 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     7261 tttgtggaac agcacaaaca ctacctggac gagatcatcg agcagatcag cgagttctcc
     7321 aagagagtga tcctggccga cgctaatctg gacaaggtgc tgagcgccta caacaagcac
     7381 agagacaagc ctatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     7441 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     7501 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     7561 acacggatcg acctgtctca gctgggaggc gacgcctatc cctatgacgt gcccgattat
     7621 gccagcctgg gcagcggctc ccccaagaaa aaacgcaagg tggaagatcc taagaaaaag
     7681 cggaaagtgg acggcattgg tagtgggagc aacggcagca gcggatccgg cgcaacaaac
     7741 ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgac cgagtacaag
     7801 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     7861 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     7921 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     7981 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     8041 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     8101 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     8161 gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     8221 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     8281 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     8341 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg ttaagtcgac
     8401 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     8461 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     8521 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     8581 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     8641 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     8701 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     8761 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     8821 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     8881 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     8941 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     9001 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     9061 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     9121 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     9181 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     9241 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc
     9301 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     9361 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     9421 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     9481 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     9541 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     9601 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     9661 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     9721 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     9781 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     9841 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     9901 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     9961 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
    10021 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
    10081 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
    10141 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
    10201 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
    10261 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
    10321 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
    10381 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
    10441 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
    10501 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
    10561 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
    10621 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
    10681 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
    10741 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
    10801 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
    10861 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
    10921 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
    10981 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
    11041 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
    11101 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
    11161 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
    11221 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
    11281 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
    11341 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
    11401 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    11461 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    11521 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    11581 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    11641 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    11701 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    11761 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    11821 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    11881 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    11941 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    12001 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    12061 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    12121 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    12181 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    12241 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    12301 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    12361 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    12421 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    12481 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    12541 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    12601 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    12661 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    12721 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    12781 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    12841 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    12901 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    12961 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    13021 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    13081 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    13141 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    13201 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    13261 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
    13321 gggagatctc cc
//
