LOCUS Exported 9658 bp ds-DNA circular SYN 19-APR-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS 87258 aligned to Addgene result SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9658) AUTHORS Kevin Brown TITLE Direct Submission JOURNAL Exported Wednesday, Apr 19, 2017 from SnapGene 3.3.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9658 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1..2720 /note="pTUB1 (FULL)" CDS 2730..4535 /codon_start=1 /note="OsTIR1-Linker-3FLAG" /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKILG GGGSDYKDHDGDYKDHDIDYKDDDDK" misc_feature 4548..5290 /note="DHFR 3' UTR" primer_bind complement(5304..5320) /note="SK primer" /note="common sequencing primer, one of multiple similar variants" promoter complement(5353..5371) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5378..5394) /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" rep_origin 5536..5991 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 6017..6121 /gene="bla" /note="AmpR promoter" CDS 6122..6982 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 7153..7741 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 8065..8095 /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 8103..8119 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 8127..8143 /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" promoter 8164..8182 /note="T3 promoter" /note="promoter for bacteriophage T3 RNA polymerase" primer_bind 8211..8227 /note="KS primer" /note="common sequencing primer, one of multiple similar variants" misc_feature 8263..8623 /note="pSAG1" CDS 8624..9286 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /note="CmR" /note="confers resistance to chloramphenicol" /translation="MHEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFS SLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVAN MDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" misc_feature complement(8956..8975) /note="CAT sgRNA target" misc_feature 9290..9590 /note="SAG1 3' UTR" primer_bind complement(9604..9620) /note="SK primer" /note="common sequencing primer, one of multiple similar variants" primer_bind 9628..9644 /note="KS primer" /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 cagcatcatc tctggaagca tcccctgaac tgcctgagtc taccaagagc actggcgaag 61 gctgtgagta gtcggacagg cacggtgact catgttgttg gacagtagcg agctctgggt 121 taaccgcata ttcactaact ggctccgtcc tgttgtcatt agatactgaa tcaggtaacg 181 atacatgagc agcatcctcg tgttccaggc gcatgtcctg ctccggcttg caaccaagga 241 cccgtggttc atctttgggt ctctccgtac tgggtggtag aggtgaaact gtcgacgtgg 301 atgcagctgc cctgcttaga gtacggacga agtgaatagc tgcgtctgca tgaacaaggg 361 gctctgaggc ccgctgtgat acgaaaggtt tgctggctac tgaacatagg tctcgcagtg 421 cgggggcata ctccagtcgg ccttcacgaa acttcgtgac caggcgatca acaagcggcg 481 tcacgcgagt taaatccgtc cagagaaagc caccatagtg caccatatac tgccggcaca 541 tcttgctgaa agtcgtggcg tgtcgatcag ctaagtcctg tgtgtacttg gaagtgagca 601 gctcttccag ttcggtccag tcatattgcc atgccctagc caccgtttca ccagcgtccg 661 atgcaggctc acctagccga tacctgtgac cgttcatata tgagttgcta ttacatctgt 721 cggccgtaac gacacaagga gatgcggctg gaggcaacgg gtttgtgagc accattttga 781 ctctgtcacg gagtaaaaaa catttattcg aactttgtac gagcgcagtc agtagaagtc 841 aaccacgcgt atcaactacg ctgcaattac agaggacagg gacagggaaa aaaagccgaa 901 gaggttgccg gtgttaggag atgacgagac gtttaaacgg ccgctggcta ttgttcgggc 961 atcgttgagc cacaagcgat caaggtgaaa acaaagttaa atagttatgc tggagcgatt 1021 gccctgctga atcttcagat cggacgaaca gtcattctgg cccactgtac ttgatgtgtt 1081 cgatgtaaat cacactcagt cgcgtgctcg gtagcaatca agttgctctt ttctctcctt 1141 tctagacacg gtaagaacgc ttatgaacac gcatacacgc atagtttttg ctagaatgca 1201 gcgaccagat gtcgcaaggt cgtctccccc atcgactgga gaatcaagaa aaacctgcgt 1261 tgatcccaaa cgtactctgt ggttggtgca accagaagtt tcatactgat caaaagccag 1321 tgaacagctg ggggacattg caggtctggt gcttcaagaa gcgctgaaga agaaagtggc 1381 gaaaccctcg gcagttgcct tggaagaggc gccgtgcatg taacttctga agtgcgtagt 1441 acctggctcc taatgctgtt ttgtgtttcg ctgtctgggc agcagtagaa tgctgtgcca 1501 gaattagcca ctattttaga catttattta cacatttttt ttctgatgaa cttggcttat 1561 tcattttttc aagtcttgcc actgggtggt ggcatgagac tcgcttagat gtatgtgggt 1621 gttgcaatca cgctggatgc tcggcttatt tctgagtttt ttgtggtttt gacaatggga 1681 acgatttcag agctactatt tcacgtggta cggttatgag ccactaaaaa aacgaagaaa 1741 aacgctgttt gcagaaacaa tagcaaactg tttttcgtca tagtaactca gcgccccctt 1801 gccccccccc agcagtgaga tgcaagacaa tcctctctac cacagctttt gttgcgtctg 1861 tttcaaattt tcagcgctcg cgaaaggcat cacgaacaac attatgagag ggagcaggtt 1921 tgtggggctg gcgggtgcag gaatgtgttc ctgcgaaaaa ggcctctgct gagaaggtcg 1981 tggcgtttga aaaatatccg aggtagcaaa gacttgtttc agtgctcccc ttttgaagac 2041 ctgcggcggc agtgcactga agagtaactc caaatcaccg cggtgagact tggttttttc 2101 cgttatcctt cagaagagtg tgttttcgtt taattcgtca cagaccacga aaaacgaacc 2161 atcgaagacg atcactgcgt ccgcgtgcat ctggatggat gacccacatc tgttgcagcc 2221 gtcgcagaca tgcatgtccc gcgttcgtga aattctctgc atcagcggag tgatcaggaa 2281 tcatcgtctc agcgggatga cgttgcggag caggccggct cgcggtgggc agtcagatgc 2341 cgaaggcgta actcaggacg gcttgcgctc atcgcagaac aggggtggtg cctgcattgg 2401 gtgcggttgg tgatcctggt tggaccggtg gagatgcgcg cgcacgaagg ggatgtgtca 2461 gaaacatttt gtttgttctc tgtgaacttt tagatgtgtt aaaggcggcg aatattagca 2521 gagagtcctc cttgttggat tctctcttga atttcgccct ttccttctct ttgcgagtct 2581 cgtagagaac aagcactcgt tcgccgtccc tgacgacgca acccgcgcag aagacatcca 2641 ccaaacggtg ttacacaatc accttgtgtg aagttcttgc ggaaaactac tcgttggcat 2701 tttttcttga attccctttt agatctaaaa tgacctactt ccctgaggaa gtggtcgagc 2761 acattttctc tttcctcccc gcacaaaggg acaggaacac ggttagtctg gtctgcaagg 2821 tgtggtacga gatcgaacgc cttagccgcc gaggtgtttt cgtcggaaat tgctacgctg 2881 tgcgtgccgg tcgggtcgcc gcccgatttc ccaatgtccg agcgctcacc gtgaaaggaa 2941 agccacactt tgcggatttt aatctcgtgc ctcccgactg gggtgggtac gctggcccct 3001 ggatcgaagc tgcagcgagg ggatgtcacg gcctcgaaga actgcgtatg aaaagaatgg 3061 tcgttagcga cgaaagcctg gaacttttgg cacgttcgtt tccacgcttc cgcgcgctgg 3121 tccttatctc ctgcgaagga ttctctacgg acggtttggc agcggtcgcc tctcattgta 3181 aactcttgcg agagctcgat ctgcaggaga acgaggtgga ggatcgagga cctcgctggc 3241 tgtcttgctt cccggacagt tgtactagcc ttgtgagcct gaacttcgca tgtattaagg 3301 gtgaagtgaa cgccggctcc ctggagcgct tggtgtcccg ctcccccaac cttcgttctc 3361 tcagactgaa cagaagcgtc agcgttgaca cccttgccaa gatcctgctg cgaacgccga 3421 acctcgagga cttgggcaca ggcaatctca ctgatgactt ccaaaccgag tcgtatttca 3481 agctgacatc tgctttggaa aagtgtaaga tgctccggag cctgagtggc ttttgggatg 3541 caagcccggt ctgcctctcg ttcatctacc ccttgtgtgc gcaactgaca ggcctcaatc 3601 tttcgtatgc cccgacattg gatgcctctg acctcaccaa aatgatctcc agatgtgtta 3661 agctccaacg gctctgggtg cttgattgca ttagcgacaa gggcctgcaa gtggtcgcgt 3721 cctcttgcaa ggatctgcag gagctgcgcg tgtttccatc tgacttctat gtcgccggat 3781 atagtgcagt cacggaagag ggactcgtgg cggtctctct gggctgccct aagcttaact 3841 ctctcctcta cttctgccat cagatgacca acgccgcgct cgtgacggtc gcgaagaact 3901 gccccaactt cacccgcttt agactctgta tccttgaacc agggaagccc gacgtggtca 3961 cgtcgcagcc tctcgatgag ggctttggcg cgatcgttcg ggaatgcaag ggtttgcaac 4021 gtttgtcgat tagcgggctt ttgacggata aggtttttat gtacatcgga aagtacgcca 4081 agcagctcga aatgctgtcc attgctttcg ccggggacag cgacaagggg atgatgcatg 4141 ttatgaatgg atgcaagaac ctgagaaagc tggagatccg cgattcgccg ttcggcgatg 4201 cggccctcct gggaaacttc gcccgatacg agactatgcg ttctctctgg atgtcgagct 4261 gtaacgtcac gcttaaaggt tgccaagttc ttgcaagcaa gatgcccatg ttgaacgtgg 4321 aagtgattaa cgagagggac ggatctaacg agatggagga gaaccacggc gacctcccaa 4381 aggtggaaaa gctttacgtc tacaggacaa ctgcaggagc gagagacgat gccccaaact 4441 ttgtcaaaat cctgggtgga ggtggaagtg actataagga ccacgacgga gactacaagg 4501 atcatgatat tgattacaaa gacgatgacg ataagtaaat gcatgccaca gaagctgccc 4561 gtctctcgtt ttcctctctt ttcggaggga tcagggagag tgcctcgggt cggagagagc 4621 tgacgagggg gtgccagaga cccctgtgtc ctttatcgaa gaaaagggat gactcttcat 4681 gtggcatttc acacagtctc acctcgcctt gttttctttt tgtcaatcag aacgaaagcg 4741 agttgcgggt gacgcagatg tgcgtgtatc cactcgtgaa tgcgttatcg ttctgtatgc 4801 cgctagagtg ctggactgtt gctgtctgcc cacgacagca gacaactttc cttctatgca 4861 cttgcaggat ggtgcagcgc aaacgacgga gagaaaggag caccctctca gtttccctac 4921 gatgtgctgt cagtttcgac tcttcaccgc gaacgattgg cgatacgtct ctgttgactt 4981 gttaggctcc gaccacgaag ctcccttaac tagataagcc gcgacaccta agtgtacacc 5041 atttgcagat cgataatctg cgaccgctga atccgtccag atcagtaaaa ccgcaccacc 5101 taagtgtaaa ccttgtttag gtcgataaaa tgctaccaac ccccacccac aatcgagcct 5161 tgagcgtttc tgcgcacgcg ttggcctacg tgacttgctg atgcctgcct ctggccattc 5221 atgccagtca gtgcgcataa aaatgtggac acagtcggtt gacaagtgtt ctggcaggct 5281 acagtgacac cgcggtggag ggggatccac tagttctaga gcggccgcca ccgcggtgga 5341 gctccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc 5401 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg 5461 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc 5521 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 5581 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 5641 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 5701 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 5761 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 5821 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 5881 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 5941 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac 6001 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 6061 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 6121 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 6181 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 6241 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 6301 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 6361 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt 6421 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 6481 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 6541 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 6601 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 6661 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 6721 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 6781 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc 6841 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 6901 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 6961 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 7021 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 7081 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 7141 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 7201 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 7261 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 7321 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 7381 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 7441 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 7501 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 7561 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 7621 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 7681 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 7741 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 7801 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 7861 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga 7921 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg 7981 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta 8041 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg 8101 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct 8161 cgaaattaac cctcactaaa gggaacaaaa gctggtaccg ggccccccct cgaggtcgac 8221 ggtatcgata agcttgatat cgaattcctg cagcccccga gacgcgtgtt ctaaccacaa 8281 accttgagac gcgtgttcca accacgcacc ctgacacgcg tgttccaacc acgcaccctg 8341 agacgcgtgt tctaaccacg caccctgaga cgcgtgttct aaccacgcac cctgagacgc 8401 gtgttctgcc gcacaatgtg cacctgtagg aagctgtagt cactgctgat tctcactgtt 8461 ctcggcaagg gccgacgacc ggagtacagt ttttgtgggc agagccgttg tgcagctttc 8521 cgttcttctc ggttgtgtca catgtgtcat tgtcgtgtaa acacacggtt gtatgtcggt 8581 ttcgctgcac cacttcatta tttcttctgg ttttttgacg agtatgcatg agaaaaaaat 8641 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt 8701 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt 8761 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg 8821 cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg 8881 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct 8941 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc 9001 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt 9061 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa 9121 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat 9181 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct 9241 taatgaatta caacagtact gcgatgagtg gcagggcggg gcttaattaa tcaccgttgt 9301 gctcacttct caaatcgaca aaggaaacac acttcgtgca gcatgtgccc cattataaag 9361 aaactgagtt gttccgttgt ggcttgcagg tgtcacatcc acaaaaaccg gccgactcta 9421 aataggagtg tttcgcagca agcagcgaaa gtttatgact gggtccgaat ctctgaacgg 9481 atgtgtggcg gacctggctg atgttgatcg ccgtcgacac acgcgccaca tgggtcaata 9541 cacaagacag ctatcagttg ttttagtcga accggttaac acaattcttg cccccccgag 9601 ggggatccac tagttctaga gcggcctcga ggtcgacggt atcgataagc tagagctt //