LOCUS       sequence-323774-       10381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ciCas9 and mRFP in mammalian cells (an insertion vector to
            generate ciCas9-knockin cell lines).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10381)
  AUTHORS   Park J, Lim JM, Jung I, Heo SJ, Park J, Chang Y, Kim HK, Jung D, Yu 
            JH, Min S, Yoon S, Cho SR, Park T, Kim HH
  TITLE     Recording of elapsed time and temporal information about biological 
            events using Cas9.
  JOURNAL   Cell. 2021 Jan 28. pii: S0092-8674(21)00014-3. doi: 
            10.1016/j.cell.2021.01.014.
  PUBMED    33539780
REFERENCE   2  (bases 1 to 10381)
  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: "Cell. 2021 
            Jan 28. pii: S0092-8674(21)00014-3. doi: 
            10.1016/j.cell.2021.01.014."
COMMENT     Sequence Label: pcDNA5/FRT/CMV_promoter-ciCas9-E2A-mRFP
FEATURES             Location/Qualifiers
     source          1..10381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      738..747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             747..770
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2748..5132
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="CFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILED
                     IVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK
                     TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKG
                     ILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQI
                     LKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDN
                     KVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDK
                     AGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF
                     YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGK
                     ATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMP
                     QVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKV
                     EKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELEN
                     GRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLD
                     EIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKY
                     FDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
     CDS             5145..5165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5277..5336
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             5343..6056
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     primer_bind     complement(6099..6116)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6105..6329
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6462..6481)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    6613..6660
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             6668..7690
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             6668..7690
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    7820..7941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7857..7876)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7911..7930
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7990..8006)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7990..8006)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8003..8025)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8014..8030
                     /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(8038..8044,8045..8062,8063..8068))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8083..8104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8221..8238)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8392..8980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8472..8491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9151..9942,9943..10011))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRHIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9774..9793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10012..10116)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10191..10210)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttgcc accatggact acaaggacga cgacgacaag gacaagaagt
      781 actccattgg gctcgatatc ggcacaaaca gcgtcggctg ggccgtcatt acggacgagt
      841 acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac agcataaaga
      901 agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc acgcggctca
      961 aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac ctgcaggaga
     1021 tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg gaggagtcct
     1081 ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat atcgtggacg
     1141 aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag cttgtagaca
     1201 gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg atcaaatttc
     1261 ggggacactt cctcatcgag ggggacctga acccagacaa cagcagcaac cgggagctgg
     1321 tggttgactt tctctcctac aagctttccc agaaaggata cagctggagt cagtttagtg
     1381 atgtggaaga gaacaggact gaggccccag aagggactga atcggagatg gagaccccca
     1441 gtgccatcaa tggcaaccca tcctggcacc tggcagacag ccccgcggtg aatggagcca
     1501 ctggccacag cagcagtttg gatgcccggg aggtgatccc catggcagca gtaaagcaag
     1561 cgctgaggga ggcaggcgac gagtttgaac tgcggtaccg gcgggcattc agtgacctga
     1621 catcccagct ccacatcacc ccagggacag catatcagag ctttgaacag gtagtgaatg
     1681 aactcttccg ggatggggta aactggggtc gcattgtggc ctttttctcc ttcggcgggg
     1741 cactgtgcgt ggaaagcgta gacaaggaga tgcaggtatt ggtgagtcgg atcgcagctt
     1801 ggatggccac ttacctgaat gaccacctag agccttggat ccaggagaac ggcggctggg
     1861 atacttttgt ggaactctat gggaacaatg gtagcggaac tgcttctggg actgggagtg
     1921 gcaccggttc agcaacagga agcggcactg tgaacacgga gatcaccaaa gctccgctga
     1981 gcgctagtat gatcaagcgc tatgatgagc accaccaaga cttgactttg ctgaaggccc
     2041 ttgtcagaca gcaactgcct gagaagtaca aggaaatttt cttcgatcag tctaaaaatg
     2101 gctacgccgg atacattgac ggcggagcaa gccaggagga attttacaaa tttattaagc
     2161 ccatcttgga aaaaatggac ggcaccgagg agctgctggt aaagcttaac agagaagatc
     2221 tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc ccaccagatt cacctgggcg
     2281 aactgcacgc tatcctcagg cggcaagagg atttctaccc ctttttgaaa gataacaggg
     2341 aaaagattga gaaaatcctc acatttcgga taccctacta tgtaggcccc ctcgcccggg
     2401 gaaattccag attcgcgtgg atgactcgca aatcagaaga gaccatcact ccctggaact
     2461 tcgaggaagt cgtggataag ggggcctctg cccagtcctt catcgaaagg atgactaact
     2521 ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg tacgagtact
     2581 tcacagttta taacgagctc accaaggtca aatacgtcac agaagggatg agaaagccag
     2641 cattcctgtc tggagagcag aagaaagcta tcgtggacct cctcttcaag acgaaccgga
     2701 aagttaccgt gaaacagctc aaagaagact atttcaaaaa gattgaatgt ttcgactctg
     2761 ttgaaatcag cggagtggag gatcgcttca acgcatccct gggaacgtat cacgatctcc
     2821 tgaaaatcat taaagacaag gacttcctgg acaatgagga gaacgaggac attcttgagg
     2881 acattgtcct cacccttacg ttgtttgaag atagggagat gattgaagaa cgcttgaaaa
     2941 cttacgctca tctcttcgac gacaaagtca tgaaacagct caagaggcgc cgatatacag
     3001 gatgggggcg gctgtcaaga aaactgatca atgggatccg agacaagcag agtggaaaga
     3061 caatcctgga ttttcttaag tccgatggat ttgccaaccg gaacttcatg cagttgatcc
     3121 atgatgactc tctcaccttt aaggaggaca tccagaaagc acaagtttct ggccaggggg
     3181 acagtcttca cgagcacatc gctaatcttg caggtagccc agctatcaaa aagggaatac
     3241 tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat gggaaggcat aagcccgaga
     3301 atatcgttat cgagatggcc cgagagaacc aaactaccca gaagggacag aagaacagta
     3361 gggaaaggat gaagaggatt gaagagggta taaaagaact ggggtcccaa atccttaagg
     3421 aacacccagt tgaaaacacc cagcttcaga atgagaagct ctacctgtac tacctgcaga
     3481 acggcaggga catgtacgtg gatcaggaac tggacatcaa tcggctctcc gactacgacg
     3541 tggatcatat cgtgccccag tcttttctca aagatgattc tattgataat aaagtgttga
     3601 caagatccga taaaaataga gggaagagtg ataacgtccc ctcagaagaa gttgtcaaga
     3661 aaatgaaaaa ttattggcgg cagctgctga acgccaaact gatcacacaa cggaagttcg
     3721 ataatctgac taaggctgaa cgaggtggcc tgtctgagtt ggataaagcc ggcttcatca
     3781 aaaggcagct tgttgagaca cgccagatca ccaagcacgt ggcccaaatt ctcgattcac
     3841 gcatgaacac caagtacgat gaaaatgaca aactgattcg agaggtgaaa gttattactc
     3901 tgaagtctaa gctggtctca gatttcagaa aggactttca gttttataag gtgagagaga
     3961 tcaacaatta ccaccatgcg catgatgcct acctgaatgc agtggtaggc actgcactta
     4021 tcaaaaaata tcccaagctt gaatctgaat ttgtttacgg agactataaa gtgtacgatg
     4081 ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa ggccaccgct aagtacttct
     4141 tttacagcaa tattatgaat tttttcaaga ccgagattac actggccaat ggagagattc
     4201 ggaagcgacc acttatcgaa acaaacggag aaacaggaga aatcgtgtgg gacaagggta
     4261 gggatttcgc gacagtccgg aaggtcctgt ccatgccgca ggtgaacatc gttaaaaaga
     4321 ccgaagtaca gaccggaggc ttctccaagg aaagtatcct cccgaaaagg aacagcgaca
     4381 agctgatcgc acgcaaaaaa gattgggacc ccaagaaata cggcggattc gattctccta
     4441 cagtcgctta cagtgtactg gttgtggcca aagtggagaa agggaagtct aaaaaactca
     4501 aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg atcaagcttc gaaaaaaacc
     4561 ccatcgactt tctcgaggcg aaaggatata aagaggtcaa aaaagacctc atcattaagc
     4621 ttcccaagta ctctctcttt gagcttgaaa acggccggaa acgaatgctc gctagtgcgg
     4681 gcgagctgca gaaaggtaac gagctggcac tgccctctaa atacgttaat ttcttgtatc
     4741 tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga taatgagcag aagcagctgt
     4801 tcgtggaaca acacaaacac taccttgatg agatcatcga gcaaataagc gaattctcca
     4861 aaagagtgat cctcgccgac gctaacctcg ataaggtgct ttctgcttac aataagcaca
     4921 gggataagcc catcagggag caggcagaaa acattatcca cttgtttact ctgaccaact
     4981 tgggcgcgcc tgcagccttc aagtacttcg acaccaccat agacagaaag cggtacacct
     5041 ctacaaagga ggtcctggac gccacactga ttcatcagtc aattacgggg ctctatgaaa
     5101 caagaatcga cctctctcag ctcggtggag acagcagggc tgaccccaag aagaagagga
     5161 aggtgacagg aagcggcact gctcccccca atctctgggc agcgcagcgc tacggccgtg
     5221 agctccgaag gatggccgat gagggtgagg gttccttcaa ggaattcgga agcgggcagt
     5281 gcaccaacta cgccctgctg aagctggccg gcgacgtgga gagcaacccc ggccccggat
     5341 ccatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag ctgtacatgg
     5401 agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg
     5461 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
     5521 acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac acccagggca
     5581 tccccgactt ctttaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
     5641 acgaggacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
     5701 tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga
     5761 aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc ggcctggaag
     5821 gcagaagcga catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttcaaga
     5881 ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg
     5941 accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg
     6001 tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt aattaaggcc
     6061 gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     6121 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     6181 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     6241 tattctgggg ggtagggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     6301 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     6361 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6421 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6481 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6541 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6601 ttcacgtacc tagaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc
     6661 cttggccaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     6721 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     6781 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     6841 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     6901 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     6961 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     7021 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     7081 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     7141 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     7201 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     7261 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     7321 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     7381 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     7441 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     7501 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     7561 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     7621 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     7681 aaaggaatag cacgtactac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     7741 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     7801 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     7861 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     7921 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     7981 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     8041 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     8101 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     8161 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     8221 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     8281 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     8341 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     8401 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     8461 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     8521 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     8581 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     8641 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     8701 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     8761 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8821 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     8881 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8941 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     9001 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     9061 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     9121 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     9181 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     9241 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     9301 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     9361 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     9421 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     9481 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     9541 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     9601 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     9661 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9721 ctgagaatag tgtatgtggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9781 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9841 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9901 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9961 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    10021 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    10081 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    10141 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
    10201 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
    10261 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
    10321 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
    10381 t
//
