LOCUS       sequence-354095-       13815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing dCas9-VPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13815)
  AUTHORS   Nghi Dang , Alissa Lance-Byrne, Angela Tung, Xiaoge Guo, Ryan J 
            Cecchi, Joanna Buchthal, Alejandro Chavez, Nan Cher Yeo
  TITLE     Generation and application of a versatile CRISPR toolkit for 
            mammalian cell engineering
  JOURNAL   Synthetic Biology, Volume 6, Issue 1, 2021
REFERENCE   2  (bases 1 to 13815)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Synthetic 
            Biology, Volume 6, Issue 1, 2021"
COMMENT     Sequence Label: NAX_CAGGS_CAS9_m4_VPR_iRFP713
FEATURES             Location/Qualifiers
     source          1..13815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          149..1165
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1089..1111
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1173..1192
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1261..1270
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1267..1293,1294..1296,1297..3780,3781..3786,3787..3852,
                     3853..3855,3856..5370)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5383..5403
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5428..5577
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             5602..5622
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6052..6408
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             6427..6996
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     primer_bind     complement(7021..7040)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7065..7113
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7345..7556
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      7566..7575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7572..8522
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDSSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     polyA_signal    8526..8647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8563..8582)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8617..8636
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   9410..9444
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9935..9951)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9935..9951)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9948..9970)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9959..9975
                     /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(9983..9989,9990..10007,10008..10013))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10028..10049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10166..10183)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10337..10925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10417..10436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11096..11887,11888..11956))
                     /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     11719..11738
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11957..12061)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(12087..12542)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12224..12245)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12436..12455
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12669..12691
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12683..12700
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12684..12700
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   12830..12892
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        13313..13692
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc
       61 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
      121 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
      181 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
      241 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
      301 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
      361 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
      421 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt ttgcgctccg
      481 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
      541 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
      601 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
      661 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
      721 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga
      781 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
      841 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
      901 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
      961 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
     1021 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     1081 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     1141 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
     1201 tcattttggc aaagaatcta gataccctcg taaaggtcta gagctagcga attcgaattt
     1261 gccaccatgg acaagaagta ctccattggg ctcgctatcg gcacaaacag cgtcggctgg
     1321 gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc
     1381 gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg
     1441 gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg
     1501 atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc
     1561 cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc
     1621 tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg
     1681 aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg
     1741 gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac
     1801 agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag
     1861 aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa
     1921 tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt
     1981 ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg
     2041 gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg
     2101 ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac
     2161 gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc
     2221 gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt
     2281 gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc
     2341 tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc
     2401 atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg
     2461 ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa
     2521 ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa
     2581 aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga
     2641 aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc
     2701 gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt
     2761 gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc
     2821 acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca
     2881 ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa
     2941 gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt
     3001 gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg
     3061 aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac
     3121 attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact
     3181 tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga
     3241 tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca
     3301 atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat
     3361 gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac
     3421 agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg
     3481 cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat
     3541 atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg
     3601 gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa
     3661 cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac
     3721 ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg
     3781 gctgctatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca
     3841 agatccgata aagctagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa
     3901 atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat
     3961 aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa
     4021 aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc
     4081 atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg
     4141 aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc
     4201 aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc
     4261 aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt
     4321 aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt
     4381 tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg
     4441 aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg
     4501 gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc
     4561 gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag
     4621 ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca
     4681 gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa
     4741 agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc
     4801 atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt
     4861 cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc
     4921 gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg
     4981 gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc
     5041 gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa
     5101 agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg
     5161 gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg
     5221 ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct
     5281 acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca
     5341 agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa gaagaggaag
     5401 gtggaggcca gcggttccgg acgggctgac gcattggacg attttgatct ggatatgctg
     5461 ggaagtgacg ccctcgatga ttttgacctt gacatgcttg gttcggatgc ccttgatgac
     5521 tttgacctcg acatgctcgg cagtgacgcc cttgatgatt tcgacctgga catgctgatt
     5581 aactctagaa gttccggatc tccgaaaaag aaacgcaaag ttggtagcca gtacctgccc
     5641 gacaccgacg accggcaccg gatcgaggaa aagcggaagc ggacctacga gacattcaag
     5701 agcatcatga agaagtcccc cttcagcggc cccaccgacc ctagacctcc acctagaaga
     5761 atcgccgtgc ccagcagatc cagcgccagc gtgccaaaac ctgcccccca gccttacccc
     5821 ttcaccagca gcctgagcac catcaactac gacgagttcc ctaccatggt gttccccagc
     5881 ggccagatct ctcaggcctc tgctctggct ccagcccctc ctcaggtgct gcctcaggct
     5941 cctgctcctg caccagctcc agccatggtg tctgcactgg ctcaggcacc agcacccgtg
     6001 cctgtgctgg ctcctggacc tccacaggct gtggctccac cagcccctaa acctacacag
     6061 gccggcgagg gcacactgtc tgaagctctg ctgcagctgc agttcgacga cgaggatctg
     6121 ggagccctgc tgggaaacag caccgatcct gccgtgttca ccgacctggc cagcgtggac
     6181 aacagcgagt tccagcagct gctgaaccag ggcatccctg tggcccctca caccaccgag
     6241 cccatgctga tggaataccc cgaggccatc acccggctcg tgacaggcgc tcagaggcct
     6301 cctgatccag ctcctgcccc tctgggagca ccaggcctgc ctaatggact gctgtctggc
     6361 gacgaggact tcagctctat cgccgacatg gacttctccg cactgctggg tagcggatcg
     6421 ggatctcggg attccaggga agggatgttt ttgccgaagc ctgaggccgg ctccgctatt
     6481 agtgacgtgt ttgagggccg cgaggtgtgc cagccaaaac gaatccggcc atttcatcct
     6541 ccaggaagtc catgggccaa ccgcccactc cccgccagcc tcgcaccaac accaaccggt
     6601 ccagtacatg agccagtcgg gtcactgacc ccggcaccag tccctcagcc actggatcca
     6661 gcgcccgcag tgactcccga ggccagtcac ctgttggagg atcccgatga agagacgagc
     6721 caggctgtca aagcccttcg ggagatggcc gatactgtga ttccccagaa ggaagaggct
     6781 gcaatctgtg gccaaatgga cctttcccat ccgcccccaa ggggccatct ggatgagctg
     6841 acaaccacac ttgagtccat gaccgaggat ctgaacctgg actcacccct gaccccggaa
     6901 ttgaacgaga ttctggatac cttcctgaac gacgagtgcc tcttgcatgc catgcatatc
     6961 agcacaggac tgtccatctt cgacacatct ctgttttgag tttaaacggg ggaggctaac
     7021 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     7081 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     7141 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     7201 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     7261 cggcaggccc tgccatagca gatctgcgaa atcggatccg cggccgcact gattctagct
     7321 gcagaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     7381 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg
     7441 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     7501 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggatc
     7561 ctcgagccac catggcggaa ggatccgtcg ccaggcagcc tgacctcttg acctgcgacg
     7621 atgagccgat ccatatcccc ggtgccatcc aaccgcatgg actgctgctc gccctcgccg
     7681 ccgacatgac gatcgttgcc ggcagcgaca accttcccga actcaccgga ctggcgatcg
     7741 gcgccctgat cggccgctct gcggccgatg tcttcgactc ggaaacgcac aaccgtctga
     7801 cgatcgcctt ggccgagccc ggggcggccg tcggagcacc gatcactgtc ggcttcacga
     7861 tgcgaaagga cgcaggcttc atcggctcct ggcatcgcca tgatcagctc atcttcctcg
     7921 agctcgagcc tccccagcgg gacgtcgccg agccgcaggc gttcttccgc cgcaccaaca
     7981 gcgccatccg ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc gccgcggcgc
     8041 aagaggtgcg gaagattacc ggcttcgatc gggtgatgat ctatcgcttc gcctccgact
     8101 ccagcggcga agtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa ctaggcctgc
     8161 actatcctgc ctcaaccgtg ccggcgcagg cccgtcggct ctataccatc aacccggtac
     8221 ggatcattcc cgatatcaat tatcggccgg tgccggtcac cccagacctc aatccggtca
     8281 ccgggcggcc gattgatctt agcttcgcca tcctgcgcag cgtatcgccc gtccatctgg
     8341 agttcatgcg caacataggc atgcacggca cgatgtcgat ctcgattttg cgcggcgagc
     8401 gactgtgggg attgatcgtt tgccatcacc gaacgccgta ctacgtcgat ctcgatggcc
     8461 gccaagcctg cgagctagtc gcccaggttc tggcctggca gatcggcgtg atggaagagt
     8521 gagttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     8581 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     8641 tatcttatca tgtctggaat tgactcaaat gatgtcaatt agtctatcag aagctcatct
     8701 ggtctccctt ccgggggaca agacatccct gtttaatatt taaacagcag tgttcccaaa
     8761 ctgggttctt atatcccttg ctctggtcaa ccaggttgca gggtttcctg tcctcacagg
     8821 aacgaagtcc ctaaagaaac agtggcagcc aggtttagcc ccggaattga ctggattcct
     8881 tttttagggc ccattggtat ggctttttcc ccgtatcccc ccaggtgtct gcaggctcaa
     8941 agagcagcga gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct
     9001 gtccccgcac gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg
     9061 ggcggctcgc tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg
     9121 gggggggctg tccctgatat ctataacaag aaaatatata tataataagt tatcacgtaa
     9181 gtagaacatg aaataacaat ataattatcg tatgagttaa atcttaaaag tcacgtaaaa
     9241 gataatcatg cgtcattttg actcacgcgg tcgttatagt tcaaaatcag tgacacttac
     9301 cgcattgaca agcacgcctc acgggagctc caagcggcga ctgagatgtc ctaaatgcac
     9361 agcgacggat tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat
     9421 tttacgcaga ctatctttct agggttaatc tagctgcatc aggatcatat cgtcgggtct
     9481 tttttccggc tcagtcatcg cccaagctgg cgctatctgg gcatcgggga ggaagaagcc
     9541 cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt ttgccgagga tgactgctgc
     9601 tgcattgacg ttgagcgaaa acgcacgttt accatgatga ttcgggaagg tgtggccatg
     9661 cacgccttta acggtgaact gttcgttcag gccacctggg ataccagttc gtcgcggctt
     9721 ttccggacac agttccggat ggtcagcccg aagcgcatca gcaacccgaa caataccggc
     9781 gacagccgga actgccgtgc cggtgtgcag attaatgaca gcggtgcggc gctgggatat
     9841 tacgtcagcg aggacgggta tcctggctgg atgccgcaga aatggacatg gataccccgt
     9901 gagttacccg gcgggcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     9961 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
    10021 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
    10081 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
    10141 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
    10201 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
    10261 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
    10321 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
    10381 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
    10441 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
    10501 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
    10561 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
    10621 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
    10681 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
    10741 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
    10801 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
    10861 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
    10921 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
    10981 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
    11041 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
    11101 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
    11161 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
    11221 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
    11281 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
    11341 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
    11401 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
    11461 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    11521 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    11581 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    11641 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    11701 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    11761 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    11821 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    11881 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    11941 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    12001 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
    12061 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
    12121 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
    12181 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
    12241 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
    12301 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
    12361 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
    12421 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
    12481 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
    12541 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
    12601 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
    12661 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcct cgttcattca
    12721 cgtttttgaa cccgtggagg acgggcagac tcgcggtgca aatgtgtttt acagcgtgat
    12781 ggagcagatg aagatgctcg acacgctgca gaacacgcag ctagattaac cctagaaaga
    12841 taatcatatt gtgacgtacg ttaaagataa tcatgtgtaa aattgacgca tgtgttttat
    12901 cggtctgtat atcgaggttt atttattaat ttgaatagat attaagtttt attatattta
    12961 cacttacata ctaataataa attcaacaaa caatttattt atgtttattt atttattaaa
    13021 aaaaacaaaa actcaaaatt tcttctataa agtaacaaaa cttttatgag ggacagcccc
    13081 cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc
    13141 gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg
    13201 gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc
    13261 tctttgagcc tgcagacacc tggggggata cggggaaaag gcctccacgg ccgacattga
    13321 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
    13381 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
    13441 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
    13501 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
    13561 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
    13621 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
    13681 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
    13741 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
    13801 gggggggggg ggggg
//
