LOCUS 40924_1304 9524 bp DNA circular SYN 15-JUN-2019 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9524) AUTHORS xsan TITLE Direct Submission REFERENCE 2 (bases 1 to 9524) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..9524 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 104..208 /label=AmpR promoter CDS 209..1066 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1240..1828 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(2014..2156) /label=bom /note="basis of mobility region from pBR322" CDS complement(2261..2449) /codon_start=1 /label=rop /note="Rop protein, which maintains plasmids at low copy number" /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" promoter 4010..4028 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 4117..4139 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 4159..4185 /codon_start=1 /label=9xHis /note="9xHis affinity tag" /translation="HHHHHHHHH" CDS 4198..5298 /codon_start=1 /label=MBP /note="maltose binding protein from E. coli" /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF WYAVRTAVINAASGRQTVDEALKDAQT" CDS 5353..5373 /codon_start=1 /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" /translation="ENLYFQS" CDS 5380..7302 /codon_start=1 /label=Cas8 /note="accession number: WP_001194012.1" /translation="MQTLKELIASNPDDLTTELKRAFRPLTPHIAIDGNELDALTILVN LTDKTDDQKDLLDRAKCKQKLRDEKWWASCINCVNYRQSHNPKFPDIRSEGVIRTQALG ELPSFLLSSSKIPPYHWSYSHDSKYVNKSAFLTNEFCWDGEISCLGELLKDADHPLWNT LKKLGCSQKTCKAMAKQLADITLTTINVTLAPNYLTQISLPDSDTSYISLSPVASLSMQ SHFHQRLQDENRHSAITRFSRTTNMGVTAMTCGGAFRMLKSGAKFSSPPHHRLNSKRSW LTSEHVQSLKQYQRLNKSLIPENSRIALRRKYKIELQNMVRSWFAMQDHTLDSNILIQH LNHDLSYLGATKRFAYDPAMTKLFTELLKRELSNSINNGEQHTNGSFLVLPNIRVCGAT ALSSPVTVGIPSLTAFFGFVHAFERNINRTTSSFRVESFAICVHQLHVEKRGLTAEFVE KGDGTISAPATRDDWQCDVVFSLILNTNFAQHIDQDTLVTSLPKRLARGSAKIAIDDFK HINSFSTLETAIESLPIEAGRWLSLYAQSNNNLSDLLAAMTEDHQLMASCVGYHLLEEP KDKPNSLRGYKHAIAECIIGLINSITFSSETDPNTIFWSLKNYQNYLVVQPRSINDETT DKSSL" CDS 7274..8332 /codon_start=1 /label=Cas7 /note="accession number: WP_000775488.1" /translation="MKLPTNLAYERSIDPSDVCFFVVWPDDRKTPLTYNSRTLLGQMEA ASLAYDVSGQPIKSATAEALAQGNPHQVDFCHVPYGASHIECSFSVSFSSELRQPYKCN SSKVKQTLVQLVELYETKIGWTELATRYLMNICNGKWLWKNTRKAYCWNIVLTPWPWNG EKVGFEDIRTNYTSRQDFKNNKNWSAIVEMIKTAFSSTDGLAIFEVRATLHLPTNAMVR PSQVFTEKESGSKSKSKTQNSRVFQSTTIDGERSPILGAFKTGAAIATIDDWYPEATEP LRVGRFGVHREDVTCYRHPSTGKDFFSILQQAEHYIEVLSANKTPAQETINDMHFLMAN LIKGGMFQHKGD" CDS 8566..8934 /codon_start=1 /product="type I-F CRISPR-associated endoribonuclease Cas6/Csy4 [Vibrio cholerae]" /label=Cas6 /note="accession number: WP_000872476.1" /translation="MEQLQYFHISNTVLVPEDCTYVSFRRCQSIDKLTAAGLARKIRRL EKRALSRGEQFDPSSFAQKEHTAIAHYHSLGESSKQTNRNFRLNIRMLSEQPREGNSIF SSYGLSNSENSFQPVPLI" terminator 9074..9121 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(9487..9515) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" ORIGIN 1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 181 gcttcaataa cattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact 781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc 2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc 2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga 2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc 2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg 2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca 2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac 2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg 2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga 2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga 2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc 2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg 2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga 2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg 3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc 3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg 3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt 3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt 3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg 3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat 3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc 3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt 3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat 3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg 3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg 3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc 3721 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc 3781 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc 3841 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc 3901 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc 3961 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc 4021 actataggga gaccacaacg gtttccctct agtgccggct ccggagagct ctttaattaa 4081 gcggccgccc tgcaggactc gagttctaga aataattttg tttaacttta agaaggagat 4141 atacatatga aatcttctca ccatcaccat caccatcacc atcaccatgg ttcttctatg 4201 aaaatcgaag aaggtaaact ggtaatctgg attaacggcg ataaaggcta taacggtctc 4261 gctgaagtcg gtaagaaatt cgagaaagat accggaatta aagtcaccgt tgagcatccg 4321 gataaactgg aagagaaatt cccacaggtt gcggcaactg gcgatggccc tgacattatc 4381 ttctgggcac acgaccgctt tggtggctac gctcaatctg gcctgttggc tgaaatcacc 4441 ccggacaaag cgttccagga caagctgtat ccgtttacct gggatgccgt acgttacaac 4501 ggcaagctga ttgcttaccc gatcgctgtt gaagcgttat cgctgattta taacaaagat 4561 ctgctgccga acccgccaaa aacctgggaa gagatcccgg cgctggataa agaactgaaa 4621 gcgaaaggta agagcgcgct gatgttcaac ctgcaagaac cgtacttcac ctggccgctg 4681 attgctgctg acgggggtta tgcgttcaag tatgaaaacg gcaagtacga cattaaagac 4741 gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct tcctggttga cctgattaaa 4801 aacaaacaca tgaatgcaga caccgattac tccatcgcag aagctgcctt taataaaggc 4861 gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca acatcgacac cagcaaagtg 4921 aattatggtg taacggtact gccgaccttc aagggtcaac catccaaacc gttcgttggc 4981 gtgctgagcg caggtattaa cgccgccagt ccgaacaaag agctggcaaa agagttcctc 5041 gaaaactatc tgctgactga tgaaggtctg gaagcggtta ataaagacaa accgctgggt 5101 gccgtagcgc tgaagtctta cgaggaagag ttggcgaaag atccacgtat tgccgccact 5161 atggaaaacg cccagaaagg tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg 5221 tatgccgtgc gtactgcggt gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc 5281 ctgaaagacg cgcagactaa ttcgagctcg aacaacaaca acaataacaa taacaacaac 5341 ctcgggatcg aggaaaacct gtacttccaa tccaatgcaa tgcaaactct gaaagaacta 5401 atcgcatcca atcctgacga cttaacaact gaacttaaga gagcatttcg tccactcaca 5461 ccgcatattg caattgatgg taatgaactt gacgcactga cgatattagt caatttaacc 5521 gataagactg atgatcagaa agacctgctc gatcgagcca aatgcaagca aaaacttcga 5581 gatgaaaaat ggtgggctag ctgcataaat tgcgttaatt acagacaaag ccataaccca 5641 aaattcccgg atatacgttc tgaaggcgtg attcgaaccc aagccctggg tgaattaccg 5701 agtttcctac tctcgtcctc aaaaatccca ccataccatt ggtcatatag ccatgattca 5761 aaatacgtca acaaaagcgc attcctcacc aatgagtttt gttgggatgg tgagatctca 5821 tgtttgggtg agcttcttaa agatgcagat cacccacttt ggaatacttt gaaaaagtta 5881 ggttgttctc aaaaaacttg caaagcaatg gcaaaacaac tagctgatat tactctcacg 5941 actatcaatg tcacactcgc accaaattac ctgactcaaa tatctctccc cgatagtgat 6001 acatcctaca tttcactctc tcctgttgca tcgctatcga tgcaaagcca ctttcatcag 6061 aggctccaag atgaaaatcg gcatagcgcg ataacgcggt ttagccgaac taccaacatg 6121 ggggtcactg cgatgacatg tggcggcgca tttagaatgt taaagtctgg cgctaagttt 6181 tccagccccc ctcatcaccg attaaacagt aaacgaagtt ggttgacgtc agagcatgtt 6241 cagtcattaa aacagtacca gcgcctcaac aaaagcctca tacctgaaaa ctctcggatt 6301 gcactccgta gaaaatacaa aatcgagctt caaaatatgg tcagatcttg gtttgcaatg 6361 caagaccata cccttgattc gaacatactt atccaacatt tgaatcatga cctatcttac 6421 ttaggagcca caaaacgttt tgcatacgat ccagcgatga ccaagctctt tactgagctt 6481 ttgaaacgag agttatcaaa ttcaatcaat aatggtgagc aacacactaa tggatcgttt 6541 ttagtcctac cgaatatcag ggtttgtggc gcaacagctt taagctcccc ggtaacggtg 6601 gggattccat cacttacagc tttctttggc ttcgttcacg catttgaacg gaatataaat 6661 cgcaccacct catcgtttcg tgttgaatcc tttgcgatat gcgtccatca actacatgtc 6721 gagaagcgag gtttgacagc agagtttgtg gaaaaaggcg acgggactat atccgctccc 6781 gcgacccggg atgactggca gtgtgatgtc gtatttagcc ttattttgaa caccaacttt 6841 gctcaacata ttgaccaaga tacgttagtt acatcactac caaagcgatt ggctcggggt 6901 tcagcaaaaa ttgcgattga tgactttaaa catatcaact cattctcgac attagaaaca 6961 gcgatcgaat ctctgccaat agaagctggt aggtggttat cactttacgc acagtcaaac 7021 aataatctaa gtgatctatt agcagccatg acagaggacc atcagctcat ggcaagctgc 7081 gtcggttacc acttgttaga agagcccaaa gataaaccaa actccctcag aggttacaaa 7141 cacgctatcg ccgagtgcat cattggactc attaactcaa tcacctttag ctcagagact 7201 gatcccaaca caatcttttg gtcgctaaag aactatcaaa actacctagt ggtacagcca 7261 aggagtatca acgatgaaac taccgacaaa tctagcctat gagcgctcta tcgacccatc 7321 agatgtctgt ttttttgtcg tctggcccga tgatagaaaa acacctttaa cctacaattc 7381 tcgtactctg ctcgggcaaa tggaagcggc atcattagcc tatgatgtct caggtcaacc 7441 aataaaaagt gccaccgctg aggcgttagc tcaagggaac cctcatcaag ttgatttctg 7501 ccacgttcca tacggtgcga gtcatattga atgcagtttc tccgtctcgt tttcttctga 7561 actacgtcaa ccatataagt gtaactcaag caaagttaaa caaacgctag tgcaattagt 7621 cgagctctac gaaacgaaaa tcggctggac tgagctagca acccgatatt tgatgaacat 7681 ttgcaacggt aaatggctgt ggaaaaatac ccgtaaagcc tattgctgga acattgtact 7741 tacaccttgg ccttggaacg gggaaaaggt tggatttgaa gatattcgta ctaactacac 7801 ctcacggcaa gactttaaaa ataataaaaa ttggtctgct atagttgaaa tgatcaaaac 7861 cgcattttct agtactgatg ggctggcgat atttgaagtc agggccacct tgcacttgcc 7921 aacgaatgct atggtgcggc caagccaagt tttcacagaa aaagaaagtg gcagtaaaag 7981 taaatctaaa actcaaaaca gtcgagtttt tcagagtaca actattgatg gtgaacgatc 8041 gccaatacta ggggccttta aaacgggagc agctattgca accattgacg actggtatcc 8101 tgaagccact gagccactaa gggtcggacg gtttggggtt catcgcgaag atgtcacttg 8161 ctaccgtcat ccgtctaccg gaaaagattt tttctcgata ttacaacaag cagagcacta 8221 tattgaagtg ttgagcgcca acaaaactcc cgctcaagaa actatcaacg acatgcactt 8281 tttaatggct aacctgatta agggtgggat gttccagcat aaaggagact gactgtgaaa 8341 tggtattata agacaatcac ctttctgcca gagttgtgca acaacgagtc actggctgca 8401 aagtgtctcc gcgttctgca tggatttaac tatcagtatg agacacgaaa tatcggcgtt 8461 tcatttccgc tttggtgtga tgcaacggtt ggaaaaaaga tttcatttgt cagcaagaac 8521 aagatagaac tcgacttact acttaaacaa cactatttcg tccaaatgga acaacttcaa 8581 tactttcata tatccaacac tgttctcgtc ccagaagatt gtacatacgt ttcctttaga 8641 cgctgtcaat ctatagataa gctcacagca gcagggctgg caaggaaaat cagacgcctg 8701 gagaaacgtg ctctatctag aggcgagcaa tttgacccat catcttttgc tcaaaaagag 8761 catactgcaa tagcgcacta ccactcactt ggggagtcca gcaaacagac gaaccgcaac 8821 tttcgactca atatcaggat gctctcggag caaccccgtg aggggaactc gatttttagt 8881 agctatggct tatcaaattc agaaaactcg tttcagcctg tacccttaat ctaatagcat 8941 tggaagtgga taacggatcc gcgatcgcgg cgcgccacct ggtggccggc cggtaccacg 9001 cgtgcgcgct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac 9061 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt 9121 gctgaaagga ggaactatat ccggatatcc acaggacggg tgtggtcgcc atgatcgcgt 9181 agtcgatagt ggctccaagt agcgaagcga gcaggactgg gcggcggcca aagcggtcgg 9241 acagtgctcc gagaacgggt gcgcatagaa attgcatcaa cgcatatagc gctagcagca 9301 cgccatagtg actggcgatg ctgtcggaat ggacgatatc ccgcaagagg cccggcagta 9361 ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga 9421 gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac 9481 taccgcatta aagcttatcg atgataagct gtcaaacatg agaa //