LOCUS Exported 5918 bp ds-DNA circular SYN 30-NOV-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGrpE-Wasabi SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5918) AUTHORS Dan Piraner TITLE Direct Submission JOURNAL Exported Wednesday, Nov 30, 2016 from SnapGene 3.2.1 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5918 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 47..168 /note="GrpE Promoter" /note="From NC_000913.3" CDS 169..171 /codon_start=1 /product="start codon" /note="ATG" /translation="M" CDS 181..198 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS 211..921 /codon_start=1 /product="green variant of teal fluorescent protein mTFP1 (Ai et al., 2008)" /note="mWasabi" /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG TNTINLEVKEGAPLPFSYDILTTAFSYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKETTGWDASTERMYVRDGVLK GDVKMKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYEIA VARNSTDGMDELYKG" terminator 1023..1070 /note="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1107..1562 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(1677..2537) /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" promoter complement(2538..2630) /gene="bla" /note="AmpR promoter" rep_origin 2711..3299 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3729..3920) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /note="rop" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4492..5574) /codon_start=1 /gene="lacI" /product="lac repressor" /note="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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(5575..5652) /gene="lacI" /note="lacI promoter" ORIGIN 1 atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg cgaaattttt ccgaggtcct 61 tgttgcgaag attgatgaca atgtgagtgc ttcccttgaa accctgaaac tgatccccat 121 aataagcgaa gttagcgaga tgaatgcgaa aaaaacgcgg agaaattcat gggcagcagc 181 catcaccatc atcaccacag ccaggatcca atggtgagca agggcgagga gaccacaatg 241 ggcgtaatca agcccgacat gaagatcaag ctgaagatgg agggcaacgt gaatggccac 301 gccttcgtga tcgagggcga gggcgagggc aagccctacg acggcaccaa caccatcaac 361 ctggaggtga aggagggagc ccccctgccc ttctcctacg acattctgac caccgcgttc 421 agttacggca acagggcctt caccaagtac cccgacgaca tccccaacta cttcaagcag 481 tccttccccg agggctactc ttgggagcgc accatgacct tcgaggacaa gggcatcgtg 541 aaggtgaagt ccgacatctc catggaggag gactccttca tctacgagat acacctcaag 601 ggcgagaact tcccccccaa cggccccgtg atgcagaagg agaccaccgg ctgggacgcc 661 tccaccgaga ggatgtacgt gcgcgacggc gtgctgaagg gcgacgtcaa gatgaagctg 721 ctgctggagg gcggcggcca ccaccgcgtt gacttcaaga ccatctacag ggccaagaag 781 gcggtgaagc tgcccgacta tcactttgtg gaccaccgca tcgagatcct gaaccacgac 841 aaggactaca acaaggtgac cgtttacgag atcgccgtgg cccgcaactc caccgacggc 901 atggacgagc tgtacaaggg ctgagtaaag aaaccgctgc tgcgaaattt gaacgccagc 961 acatggactc gtctactagc gcagcttaat taacctaggc tgctgccacc gctgagcaat 1021 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag 1081 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg 1141 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 1201 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 1261 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 1321 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 1381 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 1441 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 1501 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat 1561 ttctggcggc acgatggcat gagattatca aaaaggatct tcacctagat ccttttaaat 1621 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 1681 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 1741 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 1801 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 1861 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 1921 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 1981 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 2041 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 2101 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 2161 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 2221 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 2281 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 2341 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 2401 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 2461 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 2521 aaatgttgaa tactcatact cttccttttt caatcatgat tgaagcattt atcagggtta 2581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggtcatga 2641 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 2701 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 2761 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 2821 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag 2881 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 2941 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 3001 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 3061 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 3121 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 3181 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 3241 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 3301 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 3361 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 3421 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 3481 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatatg 3541 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta 3601 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc 3661 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc 3721 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc 3781 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg 3841 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg 3901 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta 3961 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc 4021 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga 4081 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt 4141 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc 4201 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca 4261 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa 4321 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcta 4381 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt 4441 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc 4501 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 4561 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa 4621 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt 4681 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct 4741 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc 4801 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg 4861 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc 4921 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc 4981 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg 5041 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat 5101 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc 5161 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac 5221 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac 5281 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat 5341 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt 5401 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc 5461 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt 5521 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac 5581 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca 5641 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc 5701 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga 5761 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc 5821 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc 5881 cagcaaccgc acctgtggcg ccggtgatgc cggccacg //