pCMV-PE2-P2A-GFP

LOCUS       pCMV-PE2-P2A-GFP               10569 bp ds-DNA     circular SYN 17-MAR-2020
DEFINITION  Prime editing in mammalian cells with co-translational GFP 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-PE2-P2A-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10569)
  AUTHORS   Anzalone AV, Randolph PB, Davis JR, Sousa AA, Koblan LW, Levy JM, 
            Chen PJ, Wilson C, Newby GA, Raguram A, Liu DR
  TITLE     Search-and-replace genome editing without double-strand breaks or 
            donor DNA.
  JOURNAL   Nature. 2019 Oct 21. pii: 10.1038/s41586-019-1711-4. doi: 
            10.1038/s41586-019-1711-4.
  PUBMED    31634902
REFERENCE   2  (bases 1 to 10569)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 17, 2020 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10569
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      282..870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     771..790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1024..1041
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1158..1179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1194..1224
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1232..1248
                     /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     1237..1259
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1256..1272
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1256..1272
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1343..1567
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1556..1573
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1596..1613)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1622..1642)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1655..2371)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1697..1718)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2046..2065
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2305..2326
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2372..2428)
                     /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             complement(2438..2458)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4631..8731)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(8732..8752)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8801..8820)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8802..8820)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8862..9065)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(8891..8911)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        9066..9445
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     9617..9636
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        9715..9819
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9820..111
                     /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(10038..10057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
       61 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
      121 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
      181 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
      241 cactgagtgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      421 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      901 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      961 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1021 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     1081 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     1141 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccta ggctttacac
     1201 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     1261 aacagctatg accatgatta cgccaagctc tagctagagg tcgacggtat cgagccccag
     1321 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     1381 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     1441 ctactcagac aatgcgatgc aattttctca ttttattagg aaaggacagt gggagtggca
     1501 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     1561 ggcacagtcg aggctgatca gcgggtttaa actcaatggt gatggtgatg atgaccggtt
     1621 agactttcct cttcttcttg ggagaaccac cagacttgta cagctcgtcc atgccgagag
     1681 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     1741 ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     1801 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     1861 tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga
     1921 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     1981 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     2041 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     2101 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     2161 actgcactcc ataggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     2221 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     2281 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     2341 cggtgaacag ctcctcgccc ttgctcacca taggtccagg gttctcctcc acgtctccag
     2401 cctgcttcag caggctgaag ttagtagctc cgcttccgac tttcctcttc ttcttgggct
     2461 cgaattcgct gccgtcggcg gttctttttg agccgccaga gggtgatgaa ttttctatga
     2521 ggagggtaga ggtgtctgga gtctctgtga tggctgcctt tcgggccgct tggtcagcca
     2581 tccggttgcc tctagcctcg gcgctgtgtc ccttttgatg tcctggacaa tggattatgc
     2641 taagtctttt gggcagaaag agggctttta gtagggccaa gatctcgtct ttatttttga
     2701 tctctttgcc ttctgatgtg agccacccac gccttctgta tatttctcca tggatatggg
     2761 cagtagcaaa agcataacgg ctatcagtat aaacatttag cttcttacct tctgccatct
     2821 ttagggcctg ggtgagtgct atcagttcag cccgctgagc ggatgtcccg gctggcaggg
     2881 ctttagccca gattacctcg gtctcggtgg tcaccgcagc tcccgcctta cgctgtccct
     2941 cttgtaagag actgcttcca tccgtgtacc aggtgtggtc ggcgtctggg agcggctggt
     3001 ccgttaggtc gggtcgggtt ccgtgggctt cggccaggat atcaaggcag ttgtgttgca
     3061 gcccttcctc aggcagtggg agcagcgtag ccgggttcag ggctaccacc ggtccgaact
     3121 ggacccggtc cgtgtccaaa agcaaggcct gatagtgagt catccgggcg ttggaaagcc
     3181 agcggtcggg gggttgtttg actagtgcct ctactgcatg gggggccaga atgactagtg
     3241 gctgtcccat ggttagcttg cctgcatcct ttgtcagtac ggcaatggct gctaccatcc
     3301 gtaggcaagg gggccaccca gctgctactg ggtctagctt tttggacagg taggccaccg
     3361 gccgacgcca aggtcccagt ttttgcgtta ggacaccttt ggcgtagccc tgcttctcgt
     3421 cgacaaagag ttcaaagggc ttagtcaaat ctggcaaccc cagggctggg gcagttagaa
     3481 gagcttgctt gatttcttga taggcctttt gttggtctgg gccccaatta aacagagtcc
     3541 ccggtttggt gagagggtac aggggggctg ccatttctgc aaacccaggg atgaagaggc
     3601 gacagaagcc tgccttccct aggaactccc ttagttgtcg aggggtcttc ggagtaggct
     3661 gccccatcac agtctctttt ctggcctcag tcagccatct ctgaccctct tttagaagat
     3721 accccagata cttgacctgt ttctggcaaa tttgggcttt cttggccgag gcccgatacc
     3781 cgaggttccc tagggtttgt aacagggccc gagtaccttg ttggcagtct agctcagaag
     3841 tggcggccag cagtaagtca tccacgtact gtagcaggat caagtctggg tgctggatcc
     3901 ggaagtctgc taggtctctg tgcagtgcct cattaaacag ggtgggactg tttttgaaac
     3961 cctgtgggag tctggtccag gtcaattgtc ctgagattcc catctctgga tctctccact
     4021 caaaggcgaa gagaggctga ctggtggggt ggagtctcag gcagaaaaag gcatccttta
     4081 aatcaagcac agtgtaccac tggtgggacg gtgggagccc gctcaagagg ttgtaagggt
     4141 tgggcacggt ggggtggatg tcttccaccc gcttgttgac ttctctcaga tcctggacag
     4201 gcctataatc attagtccct ggtttcttaa cgggtagcag gggcgtgttc cagggggact
     4261 ggcagggtac cagtattccc tggtccaaca gtctctgtat gtggggcttg atccccagtc
     4321 tggcttcttg tgacatgggg tattgtttta tggacacggg ggtagaggtt gctttcagag
     4381 gtatgatcag aggagcttgg cgaactgcca gtcccatgcc cccggtttcc gcccaggcct
     4441 gaggaaaatc agacagccat gtggacccta gagaaacatc tggctctttt gaggtctcat
     4501 gtagccgata ctcatcttct atatttaggg tgctgctgcc gccgctgctg ccgccactgc
     4561 tctctggtgt tgctgactcg cttgttcctg gtgtctcgct gccagaggat cctccgctag
     4621 atcctccaga gtcacctccc agctgagaca ggtcgatccg tgtctcgtac aggccggtga
     4681 tgctctggtg gatcagggtg gcgtccagca cctctttggt gctggtgtac ctcttccggt
     4741 cgatggtggt gtcaaagtac ttgaaggcgg caggggctcc cagattggtc agggtaaaca
     4801 ggtggatgat attctcggcc tgctctctga tgggcttatc ccggtgcttg ttgtaggcgg
     4861 acagcacttt gtccagatta gcgtcggcca ggatcactct cttggagaac tcgctgatct
     4921 gctcgatgat ctcgtccagg tagtgcttgt gctgttccac aaacagctgt ttctgctcat
     4981 tatcctcggg ggagcccttc agcttctcat agtggctggc caggtacagg aagttcacat
     5041 atttggaggg cagggccagt tcgtttccct tctgcagttc gccggcagag gccagcattc
     5101 tcttccggcc gttttccagc tcgaacaggg agtacttagg cagcttgatg atcaggtcct
     5161 ttttcacttc tttgtagccc ttggcttcca gaaagtcgat gggattcttc tcgaagctgc
     5221 ttctttccat gatggtgatc cccagcagct ctttcacact cttcagtttc ttggacttgc
     5281 ccttttccac tttggccacc accagcacag aataggccac ggtggggctg tcgaagccgc
     5341 cgtacttctt agggtcccag tccttctttc tggcgatcag cttatcgctg ttcctcttgg
     5401 gcaggataga ctctttgctg aagccgcctg tctgcacctc ggtctttttc acgatattca
     5461 cttggggcat gctcagcact ttccgcacgg tggcaaaatc ccggccctta tcccacacga
     5521 tctccccggt ttcgccgttt gtctcgatca gaggccgctt ccggatctcg ccgttggcca
     5581 gggtaatctc ggtcttgaaa aagttcatga tgttgctgta gaagaagtac ttggcggtag
     5641 ccttgccgat ttcctgctcg ctcttggcga tcatcttccg cacgtcgtac accttgtagt
     5701 cgccgtacac gaactcgctt tccagcttag ggtacttttt gatcagggcg gttcccacga
     5761 cggcgttcag gtaggcgtcg tgggcgtggt ggtagttgtt gatctcgcgc actttgtaaa
     5821 actggaaatc cttccggaaa tcggacacca gcttggactt cagggtgatc actttcactt
     5881 cccggatcag cttgtcattc tcgtcgtact tagtgttcat ccgggagtcc aggatctgtg
     5941 ccacgtgctt tgtgatctgc cgggtttcca ccagctgtct cttgatgaag ccggccttat
     6001 ccagttcgct caggccgcct ctctcggcct tggtcagatt gtcgaacttt ctctgggtaa
     6061 tcagcttggc gttcagcagc tgccgccagt agttcttcat cttcttcacg acctcttcgg
     6121 agggcacgtt gtcgctcttg ccccggttct tgtcgcttct ggtcagcacc ttgttgtcga
     6181 tggagtcgtc cttcagaaag ctctgaggca cgatagcgtc cacatcgtag tcggacagcc
     6241 ggttgatgtc cagttcctgg tccacgtaca tatcccgccc attctgcagg tagtacaggt
     6301 acagcttctc gttctgcagc tgggtgtttt ccacggggtg ttctttcagg atctggctgc
     6361 ccagctcttt gatgccctct tcgatccgct tcattctctc gcggctgttc ttctgtccct
     6421 tctgggtggt ctggttctct ctggccattt cgatcacgat gttctcgggc ttgtgccggc
     6481 ccatcacttt cacgagctcg tccaccacct tcactgtctg caggatgccc ttcttaatgg
     6541 cggggctgcc ggccagattg gcaatgtgct cgtgcaggct atcgccctgg ccggacacct
     6601 gggctttctg gatgtcctct ttaaaggtca ggctgtcgtc gtggatcagc tgcatgaagt
     6661 ttctgttggc gaagccgtcg gacttcagga aatccaggat tgtcttgccg gactgcttgt
     6721 cccggatgcc gttgatcagc ttccggctca gcctgcccca gccggtgtat ctccgccgct
     6781 tcagctgctt catcactttg tcgtcgaaca ggtgggcata ggttttcagc cgttcctcga
     6841 tcatctctct gtcctcaaac agtgtcaggg tcagcacgat atcttccaga atgtcctcgt
     6901 tttcctcatt gtccaggaag tccttgtcct tgataatttt cagcagatcg tggtatgtgc
     6961 ccagggaggc gttgaaccga tcttccacgc cggagatttc cacggagtcg aagcactcga
     7021 ttttcttgaa gtagtcctct ttcagctgct tcacggtcac tttccggttg gtcttgaaca
     7081 gcaggtccac gatggccttt ttctgctcgc cgctcaggaa ggcgggcttt ctcattccct
     7141 cggtcacgta tttcactttg gtcagctcgt tatacacggt gaagtactcg tacagcaggc
     7201 tgtgcttggg cagcaccttc tcgttgggca ggttcttatc gaagttggtc atccgctcga
     7261 tgaagctctg ggcggaagcg cccttgtcca ccacttcctc gaagttccag ggggtgatgg
     7321 tttcctcgct ctttctggtc atccaggcga atctgctgtt tcccctggcc agagggccca
     7381 cgtagtaggg gatgcggaag gtcaggatct tctcgatctt ttcccggttg tccttcagga
     7441 atgggtaaaa atcttcctgc cgccgcagaa tggcgtgcag ctctcccagg tggatctggt
     7501 gggggatgct gccgttgtcg aaggtccgct gcttccgcag caggtcctct ctgttcagct
     7561 tcacgagcag ttcctcggtg ccgtccatct tttccaggat gggcttgatg aacttgtaga
     7621 actcttcctg gctggctccg ccgtcaatgt agccggcgta gccgttcttg ctctggtcga
     7681 agaaaatctc tttgtacttc tcaggcagct gctgccgcac gagagctttc agcagggtca
     7741 ggtcctggtg gtgctcgtcg tatctcttga tcatagaggc gctcaggggg gccttggtga
     7801 tctcggtgtt cactctcagg atgtcgctca gcaggatggc gtcggacagg ttcttggcgg
     7861 ccagaaacag gtcggcgtac tggtcgccga tctgggccag caggttgtcc aggtcgtcgt
     7921 cgtaggtgtc cttgctcagc tgcagtttgg catcctcggc caggtcgaag ttgctcttga
     7981 agttgggggt caggcccagg ctcagggcaa tcaggtttcc gaacaggcca ttcttcttct
     8041 cgccgggcag ctgggcgatc agattttcca gccgtctgct cttgctcagt ctggcagaca
     8101 ggatggcctt ggcgtccacg ccgctggcgt tgatggggtt ttcctcgaac agctggttgt
     8161 aggtctgcac cagctggatg aacagcttgt ccacgtcgct gttgtcgggg ttcaggtcgc
     8221 cctcgatcag gaagtggccc cggaacttga tcatgtgggc cagggccaga tagatcagcc
     8281 gcaggtcggc cttgtcggtg ctgtccacca gtttctttct caggtggtag atggtggggt
     8341 acttctcgtg gtaggccacc tcgtccacga tgttgccgaa gatggggtgc cgctcgtgct
     8401 tcttatcctc ttccaccagg aaggactctt ccagtctgtg gaagaagctg tcgtccacct
     8461 tggccatctc gttgctgaag atctcttgca gatagcagat ccggttcttc cgtctggtgt
     8521 atcttcttct ggcggttctc ttcagccggg tggcctcggc tgtttcgccg ctgtcgaaca
     8581 gcagggctcc gatcaggttc ttcttgatgc tgtgccggtc ggtgttgccc agcaccttga
     8641 atttcttgct gggcaccttg tactcgtcgg tgatcacggc ccagcccaca gagttggtgc
     8701 cgatgtccag gccgatgctg tacttcttgt cgactttccg cttcttcttt ggtgactcga
     8761 actcgcttcc gtcggctgtc cgtttcatgg tggcggctct ccctatagtg agtcgtatta
     8821 gcggccgcgg atctctagcg gatctgacgg ttcactaaac cagctctgct tatatagacc
     8881 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt
     8941 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga
     9001 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc
     9061 atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     9121 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     9181 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     9241 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     9301 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     9361 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     9421 taataactag tcaataatca atgtcaacgc gtatatctgg cccgtacatc gcgaagcagc
     9481 gcaaaacgcc taaccctaag cagattcttc atgcaattgt cggtcaagcc ttgccttgtt
     9541 gtagcttaaa ttttgctcgc gcactactca gcgacctcca acacacaagc agggagcaga
     9601 tactggctta actatgcggc atcagagcag attgtactga gagtagaccc taggggatcg
     9661 ggagatcgat ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     9721 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     9781 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     9841 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     9901 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     9961 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
    10021 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
    10081 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
    10141 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
    10201 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
    10261 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
    10321 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
    10381 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
    10441 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
    10501 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
    10561 gggccagat
//