LOCUS       sequence-398276-       13332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a CRISPRi guide targeting POLR2D.
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-sgCiPOLR2D
FEATURES             Location/Qualifiers
     source          1..13332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(112..132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        117..446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      297..432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     359..378
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        494..541
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             560..934
                     /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    1064..1185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1101..1120)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1155..1174
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1234..1250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1234..1250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1247..1269)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1258..1274
                     /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(1282..1288,1289..1306,1307..1312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1327..1348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1465..1482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1636..2224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1716..1735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2395..3186,3187..3255))
                     /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     3018..3037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3256..3360)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3435..3454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3626..4005
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4007..4205
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4160..4180
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4223..4403
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4450..4575
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5068..5301
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5486..5530
                     /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    5828..5945
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5996..6244
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5996..6016
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6167..6186
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6266..6341
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6403..6614
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    6640..6664
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6694..6879
                     /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(6901..6927,6928..6930,6931..9417,9418..9420,
                     9421..11004)
                     /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             6901..11004
                     /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             11008..11034
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11014..11040
                     /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             11053..11073
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11080..11100
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11143..11199
                     /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             11200..11796
                     /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     11693..11713
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11812..12400
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11865..11885)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12283..12294)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12472..12705
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12731..12748)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12737..12961
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(13007..13332,1..103)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13094..13113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13304..13325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
       61 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
      121 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      181 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
      241 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
      301 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
      361 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
      421 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
      481 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
      541 caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
      601 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
      661 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
      721 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
      781 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
      841 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
      901 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
      961 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     1021 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     1081 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1141 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     1201 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1261 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1321 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1381 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1441 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1501 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1561 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1621 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1681 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1741 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1801 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1861 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1921 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1981 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2041 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2101 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2161 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2221 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2281 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2341 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2401 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2461 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2521 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2581 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2641 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2701 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2761 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2821 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2881 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2941 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3001 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3061 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3121 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3181 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3241 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3301 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3361 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     3421 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     3481 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     3541 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     3601 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     3661 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     3721 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     3781 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     3841 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     3901 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     3961 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     4021 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     4081 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     4141 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     4201 agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     4261 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     4321 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     4381 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     4441 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     4501 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     4561 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     4621 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     4681 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     4741 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     4801 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     4861 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     4921 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     4981 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     5041 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     5101 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     5161 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     5221 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     5281 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     5341 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     5401 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     5461 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     5521 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     5581 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     5641 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     5701 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     5761 tcgattagtg aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca
     5821 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     5881 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     5941 attttcgggt ttattacagg gacagcagag atccagtttg gttaattaag gtaccgaggg
     6001 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
     6061 tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
     6121 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct
     6181 taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa
     6241 caccggggcg ggtggcagcg atccggtttt agagctagaa atagcaagtt aaaataaggc
     6301 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa ttcgctagct
     6361 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
     6421 ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt
     6481 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     6541 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     6601 ccgccagaac acaggaccgg ttctagcatc gattgatcaa caagtttgta caaaaaagtt
     6661 ggcatggatg ctaagtcact aactgcctgg tcccggacac tggtgacctt caaggatgta
     6721 tttgtggact tcaccaggga ggagtggaag ctgctggaca ctgctcagca gatcgtgtac
     6781 agaaatgtga tgctggagaa ctataagaac ctggtttcct tgggttatca gcttactaag
     6841 ccagatgtga tcctccggtt ggagaaggga gaagagcccg gcggttccgg cggagggtcg
     6901 atggacaaga agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg
     6961 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
     7021 cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggaga aacagccgag
     7081 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
     7141 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
     7201 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
     7261 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
     7321 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
     7381 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
     7441 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
     7501 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     7561 cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggcaac
     7621 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     7681 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     7741 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     7801 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     7861 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     7921 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     7981 ggctacatcg atggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     8041 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     8101 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     8161 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     8221 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     8281 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     8341 gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     8401 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     8461 tacaacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     8521 agcggcgagc agaaaaaagc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     8581 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     8641 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     8701 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     8761 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     8821 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     8881 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     8941 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     9001 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     9061 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     9121 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     9181 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     9241 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     9301 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     9361 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggacgct
     9421 atcgtgcctc agagctttct gaaggacgac tccatcgata acaaagtgct gactcggagc
     9481 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     9541 aactactggc gccagctgct gaatgccaag ctgattaccc agaggaagtt cgacaatctg
     9601 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     9661 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     9721 actaagtacg acgagaacga caaactgatc cgggaagtga aagtgatcac cctgaagtcc
     9781 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     9841 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     9901 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     9961 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
    10021 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
    10081 cctctgatcg agacaaacgg cgaaacaggc gagatcgtgt gggataaggg ccgggacttt
    10141 gccaccgtgc ggaaagtgct gtctatgccc caagtgaata tcgtgaaaaa gaccgaggtg
    10201 cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga caagctgatc
    10261 gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc
    10321 tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg
    10381 aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac
    10441 tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag
    10501 tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg
    10561 cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc
    10621 cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa
    10681 cagcacaaac actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg
    10741 atcctggccg acgctaatct ggacaaggtg ctgagcgcct acaacaagca cagagacaag
    10801 cctatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc
    10861 cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa
    10921 gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc
    10981 gacctgtctc agctgggagg cgacgcctat ccctatgacg tgcccgatta tgccagcctg
    11041 ggcagcggct cccccaagaa aaaacgcaag gtggaagatc ctaagaaaaa gcggaaagtg
    11101 gacggcattg gtagtgggag caacggcagc agcggatccg gcgcaacaaa cttctctctg
    11161 ctgaaacaag ccggagatgt cgaagagaat cctggaccga ccgagtacaa gcccacggtg
    11221 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
    11281 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
    11341 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
    11401 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
    11461 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
    11521 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
    11581 ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
    11641 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
    11701 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
    11761 cgcacctggt gcatgacccg caagcccggt gcctgaacgc gttaagtcga caatcaacct
    11821 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
    11881 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
    11941 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
    12001 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
    12061 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
    12121 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
    12181 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
    12241 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
    12301 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
    12361 cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta agaccaatga
    12421 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
    12481 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
    12541 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
    12601 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
    12661 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc
    12721 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
    12781 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
    12841 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
    12901 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
    12961 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
    13021 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
    13081 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
    13141 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
    13201 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
    13261 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
    13321 actggaacaa ca
//
