LOCUS       sequence-354087-       12503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing dCas9-KRAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12503)
  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 12503)
  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_KRAB_iRFP713
FEATURES             Location/Qualifiers
     source          1..12503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          145..1161
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1085..1107
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1169..1188
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1258..1267
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1264..1290,1291..1293,1294..3777,3778..3783,3784..3849,
                     3850..3852,3853..5367)
                     /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             5380..5400
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5461..5655
                     /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
                     QLTKPDVILRLEKGEEPWLV"
     primer_bind     complement(5710..5729)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5754..5802
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6034..6245
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6255..6264
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6261..7211
                     /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    7215..7336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7252..7271)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7306..7325
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8099..8133
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8624..8640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8624..8640)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8637..8659)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8648..8664
                     /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(8672..8678,8679..8696,8697..8702))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8717..8738
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8855..8872)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9026..9614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9106..9125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9785..10576,10577..10645))
                     /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     10408..10427
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10646..10750)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10776..11231)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10913..10934)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11125..11144
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11358..11380
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11372..11389
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11373..11389
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11519..11581
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        12002..12381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat
       61 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
      121 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
      181 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
      241 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
      301 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg
      361 gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc
      421 gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttttgc gctccgcagt
      481 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg
      541 aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg
      601 gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
      661 ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
      721 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg
      781 cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt
      841 ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg
      901 aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc
      961 ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc
     1021 cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     1081 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     1141 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     1201 tttggcaaag aatctagata ccctcgtaaa ggtctagagc tagcggatcg aacccttgcc
     1261 accatggaca agaagtactc cattgggctc gctatcggca caaacagcgt cggctgggcc
     1321 gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg caataccgat
     1381 cgccacagca taaagaagaa cctcattggc gccctcctgt tcgactccgg ggagacggcc
     1441 gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa gaatcggatc
     1501 tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc tttcttccat
     1561 aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca cccaatcttt
     1621 ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata tcatctgagg
     1681 aagaagcttg tagacagtac tgataaggct gacttgcggt tgatctatct cgcgctggcg
     1741 catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc
     1801 gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt cgaagagaac
     1861 ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc
     1921 cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt
     1981 aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt cgacctggcc
     2041 gaagatgcca agcttcaact gagcaaagac acctacgatg atgatctcga caatctgctg
     2101 gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc
     2161 attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct
     2221 agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc
     2281 agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac
     2341 gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc
     2401 ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga agatctgttg
     2461 cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct gggcgaactg
     2521 cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag
     2581 attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc ccggggaaat
     2641 tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag
     2701 gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat
     2761 aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca
     2821 gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc
     2881 ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa ccggaaagtt
     2941 accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa
     3001 atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa
     3061 atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt
     3121 gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac
     3181 gctcatctct tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata tacaggatgg
     3241 gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg aaagacaatc
     3301 ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt gatccatgat
     3361 gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt
     3421 cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag
     3481 accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc
     3541 gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa cagtagggaa
     3601 aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac
     3661 ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc
     3721 agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta cgacgtggct
     3781 gctatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt gttgacaaga
     3841 tccgataaag ctagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg
     3901 aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat
     3961 ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt catcaaaagg
     4021 cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg
     4081 aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag
     4141 tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac
     4201 aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa
     4261 aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg
     4321 aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac
     4381 agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag
     4441 cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat
     4501 ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa
     4561 gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg
     4621 atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc
     4681 gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc
     4741 gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc
     4801 gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat taagcttccc
     4861 aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag
     4921 ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc
     4981 agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg
     5041 gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga
     5101 gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa gcacagggat
     5161 aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac caacttgggc
     5221 gcgcctgcag ccttcaagta cttcgacacc accatagaca gaaagcggta cacctctaca
     5281 aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta tgaaacaaga
     5341 atcgacctct ctcagctcgg tggagacagc agggctgacc ccaagaagaa gaggaaggtg
     5401 agtggtggag gaagtggcgg gtcagggtcg atggacgcga aatcacttac ggcatggtcg
     5461 agaacactgg ttacgttcaa ggacgtgttt gtggacttta cacgtgagga gtggaaattg
     5521 ctggatactg cgcaacaaat tgtgtatcga aatgtcatgc ttgagaatta caagaacctc
     5581 gtcagtctcg gataccagtt gacgaaaccg gatgtgatcc ttaggctcga aaagggggaa
     5641 gaaccttggc tggtatagaa gaattcgatc cctaccggtt agtaatgagt ttaaacgggg
     5701 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5761 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5821 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5881 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5941 acgtcggggc ggcaggccct gccatagcag atctgcgaaa tcggatccgc ggccgcactg
     6001 attctagctg cagaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     6061 ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg
     6121 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     6181 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     6241 cacaggatcc tcgagccacc atggcggaag gatccgtcgc caggcagcct gacctcttga
     6301 cctgcgacga tgagccgatc catatccccg gtgccatcca accgcatgga ctgctgctcg
     6361 ccctcgccgc cgacatgacg atcgttgccg gcagcgacaa ccttcccgaa ctcaccggac
     6421 tggcgatcgg cgccctgatc ggccgctctg cggccgatgt cttcgactcg gaaacgcaca
     6481 accgtctgac gatcgccttg gccgagcccg gggcggccgt cggagcaccg atcactgtcg
     6541 gcttcacgat gcgaaaggac gcaggcttca tcggctcctg gcatcgccat gatcagctca
     6601 tcttcctcga gctcgagcct ccccagcggg acgtcgccga gccgcaggcg ttcttccgcc
     6661 gcaccaacag cgccatccgc cgcctgcagg ccgccgaaac cttggaaagc gcctgcgccg
     6721 ccgcggcgca agaggtgcgg aagattaccg gcttcgatcg ggtgatgatc tatcgcttcg
     6781 cctccgactc cagcggcgaa gtgatcgcag aggatcggtg cgccgaggtc gagtcaaaac
     6841 taggcctgca ctatcctgcc tcaaccgtgc cggcgcaggc ccgtcggctc tataccatca
     6901 acccggtacg gatcattccc gatatcaatt atcggccggt gccggtcacc ccagacctca
     6961 atccggtcac cgggcggccg attgatctta gcttcgccat cctgcgcagc gtatcgcccg
     7021 tccatctgga gttcatgcgc aacataggca tgcacggcac gatgtcgatc tcgattttgc
     7081 gcggcgagcg actgtgggga ttgatcgttt gccatcaccg aacgccgtac tacgtcgatc
     7141 tcgatggccg ccaagcctgc gagctagtcg cccaggttct ggcctggcag atcggcgtga
     7201 tggaagagtg agttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     7261 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7321 tcatcaatgt atcttatcat gtctggaatt gactcaaatg atgtcaatta gtctatcaga
     7381 agctcatctg gtctcccttc cgggggacaa gacatccctg tttaatattt aaacagcagt
     7441 gttcccaaac tgggttctta tatcccttgc tctggtcaac caggttgcag ggtttcctgt
     7501 cctcacagga acgaagtccc taaagaaaca gtggcagcca ggtttagccc cggaattgac
     7561 tggattcctt ttttagggcc cattggtatg gctttttccc cgtatccccc caggtgtctg
     7621 caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt
     7681 gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc
     7741 ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg
     7801 gggctttggg ggggggctgt ccctgatatc tataacaaga aaatatatat ataataagtt
     7861 atcacgtaag tagaacatga aataacaata taattatcgt atgagttaaa tcttaaaagt
     7921 cacgtaaaag ataatcatgc gtcattttga ctcacgcggt cgttatagtt caaaatcagt
     7981 gacacttacc gcattgacaa gcacgcctca cgggagctcc aagcggcgac tgagatgtcc
     8041 taaatgcaca gcgacggatt cgcgctattt agaaagagag agcaatattt caagaatgca
     8101 tgcgtcaatt ttacgcagac tatctttcta gggttaatct agctgcatca ggatcatatc
     8161 gtcgggtctt ttttccggct cagtcatcgc ccaagctggc gctatctggg catcggggag
     8221 gaagaagccc gtgccttttc ccgcgaggtt gaagcggcat ggaaagagtt tgccgaggat
     8281 gactgctgct gcattgacgt tgagcgaaaa cgcacgttta ccatgatgat tcgggaaggt
     8341 gtggccatgc acgcctttaa cggtgaactg ttcgttcagg ccacctggga taccagttcg
     8401 tcgcggcttt tccggacaca gttccggatg gtcagcccga agcgcatcag caacccgaac
     8461 aataccggcg acagccggaa ctgccgtgcc ggtgtgcaga ttaatgacag cggtgcggcg
     8521 ctgggatatt acgtcagcga ggacgggtat cctggctgga tgccgcagaa atggacatgg
     8581 ataccccgtg agttacccgg cgggcgcgct tggcgtaatc atggtcatag ctgtttcctg
     8641 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     8701 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     8761 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     8821 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     8881 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     8941 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     9001 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     9061 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     9121 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     9181 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     9241 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     9301 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     9361 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     9421 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     9481 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     9541 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     9601 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     9661 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     9721 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     9781 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     9841 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     9901 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     9961 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
    10021 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
    10081 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
    10141 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
    10201 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
    10261 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
    10321 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
    10381 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
    10441 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
    10501 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
    10561 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
    10621 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
    10681 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
    10741 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
    10801 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
    10861 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
    10921 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
    10981 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
    11041 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
    11101 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
    11161 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
    11221 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
    11281 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
    11341 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcctc
    11401 gttcattcac gtttttgaac ccgtggagga cgggcagact cgcggtgcaa atgtgtttta
    11461 cagcgtgatg gagcagatga agatgctcga cacgctgcag aacacgcagc tagattaacc
    11521 ctagaaagat aatcatattg tgacgtacgt taaagataat catgtgtaaa attgacgcat
    11581 gtgttttatc ggtctgtata tcgaggttta tttattaatt tgaatagata ttaagtttta
    11641 ttatatttac acttacatac taataataaa ttcaacaaac aatttattta tgtttattta
    11701 tttattaaaa aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttatgagg
    11761 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
    11821 cagcgagccg cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca
    11881 gcgtgcgggg acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct
    11941 tctcgctgct ctttgagcct gcagacacct ggggggatac ggggaaaagg cctccacggc
    12001 cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
    12061 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
    12121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
    12181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
    12241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
    12301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
    12361 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
    12421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
    12481 atgggggcgg gggggggggg ggg
//
