LOCUS Exported 7433 bp ds-DNA circular SYN 27-SEP-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7433) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 27, 2017 from SnapGene 4.0.5 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7433 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 22..116 /note="lambda t0 terminator transcription terminator from phage lambda" promoter 673..775 /note="cat promoter promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 776..1435 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=cat /note="CmR confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" rep_origin 1986..2574 /label=ori /note="ori high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 2818..2895 /gene="lacI (mutant)" /label=lacI (mutant) promoter /note="lacIq promoter In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 2896..3978 /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="lacI 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 4027..4057 /note="lac promoter promoter for the E. coli lac operon" protein_bind 4065..4081 /label=lac repressor encoded by lacI binding site /bound_moiety="lac repressor encoded by lacI" /note="lac operator 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 4089..4105 /label=M13 rev /note="M13 rev common sequencing primer, one of multiple similar variants" primer_bind complement(4121..4137) /label=M13 fwd /note="M13 fwd common sequencing primer, one of multiple similar variants" promoter 4337..4381 /note="T5 promoter bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator" protein_bind 4357..4373 /label=lac repressor encoded by lacI binding site /bound_moiety="lac repressor encoded by lacI" /note="lac operator 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)." protein_bind 4389..4405 /label=lac repressor encoded by lacI binding site /bound_moiety="lac repressor encoded by lacI" /note="lac operator 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)." RBS 4424..4435 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 4448..5122 /codon_start=1 /product="monomeric photoswitchable Dronpa-Green1 fluorescent protein" /label=monomeric photoswitchable Dronpa-Green1 fluores /note="Dronpa-Green1" /translation="MVSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKV KEGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNA TNDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALS LEGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPR QAK" misc_feature 5159..5485 /label=ZapA /note="ZapA" misc_feature 5538..6686 /label=FtsZ /note="FtsZ" CDS 6723..7433 /codon_start=1 /product="photoactivatable monomeric derivative of DsRed fluorescent protein" /label=photoactivatable monomeric derivative of DsRed /note="PAmCherry mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" ORIGIN 1 aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc 61 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa 121 gctaggggaa ttcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt 181 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc 241 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc 301 atgacccggt cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg 361 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagga tccgggatgt 421 ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag ctgaacagga gggacagctg 481 atagaaacag aagccactgg agcacctcaa aaacaccatc atacactaaa tcagtaagtt 541 ggcagcatca cccgacgcac tttgcgccga ataaatacct gtgacggaag atcacttcgc 601 agaataaata aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg 661 aaaatgagac gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc 721 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga 781 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt 841 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac 901 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat 961 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct 1021 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt 1081 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca 1141 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat 1201 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa 1261 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa 1321 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg 1381 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt 1441 ttaaggcagt tattggtgcc cttaaacgcc tggttgctac gcctgaataa gtgataataa 1501 gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt 1561 cgttaaatag ccgcttatgt ctattgctgg tttaccggtt tattgactac cggaagcagt 1621 gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta 1681 ataattgacg ttatgatcat ttattctgcc tcccagagcc tgataaaaac ggttaccgga 1741 tcgagtcagg caactatgga tgaacgaaat agacagatcg cttagatagg tgcctcactg 1801 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 1861 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 1921 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 1981 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 2041 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 2101 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac 2161 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 2221 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 2281 gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca cacagcccag cttggagcga 2341 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 2401 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 2461 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 2521 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 2581 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 2641 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 2701 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 2761 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac 2821 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa 2881 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc 2941 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg 3001 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa 3061 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg 3121 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg 3181 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc 3241 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct 3301 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc 3361 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc 3421 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt 3481 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa 3541 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag 3601 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc 3661 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat 3721 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg 3781 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc 3841 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa 3901 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga 3961 ctggaaagcg ggcagtgagc gcaacgcaac taatgtaagt tagctcactc attaggcacc 4021 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca 4081 atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac 4141 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt 4201 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgccta 4261 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 4321 tcttcacctc gagaaatcat aaaaaattta tttgctttgt gagcggataa caattataat 4381 agattcaatt gtgagcggat aacaatttca cacagaattc attaaagagg agaaattaac 4441 tactagtatg gtgagtgtga ttaaaccaga catgaagatc aagctgcgta tggaaggcgc 4501 tgtaaatgga cacccgttcg cgattgaagg agttggcctt gggaagcctt tcgagggaaa 4561 acagagtatg gaccttaaag tcaaagaagg cggacctctg cctttcgcct atgacatctt 4621 gacaactgtg ttctgttacg gcaacagggt attcgccaaa tacccagaaa atatagtaga 4681 ctatttcaag cagtcgtttc ctgagggcta ctcttgggaa cgaagcatga attacgaaga 4741 cgggggcatt tgtaacgcga caaacgacat aaccctggat ggtgactgtt atatctatga 4801 aattcgattt gatggtgtga actttcctgc caatggtcca gttatgcaga agaggactgt 4861 gaaatgggag ccatccactg agaaattgta tgtgcgtgat ggagtgctga agggtgatgt 4921 taacatggct ctgtcgcttg aaggaggtgg ccattaccga tgtgacttca aaactactta 4981 taaagctaag aaggttgtcc agttgccaga ctatcacttt gtggaccacc acattgagat 5041 taaaagccac gacaaagatt acagtaatgt taatctgcat gagcacgccg aagcgcattc 5101 tgagctgccg aggcaggcca agggatccgc tggctccgct gctggttctg gcgaattctc 5161 tgcacaaccc gtcgatatcc aaatttttgg ccgttcactg cgtgtgaact gcccgcctga 5221 ccaaagggat gcgttgaatc aggcagcgga cgatctgaac caacggttgc aagatctgaa 5281 agaacgcact agagtcacaa atactgaaca gttggtcttc attgccgcat tgaatatcag 5341 ctatgagtta gcgcaagaaa aagcaaagac tcgtgactac gcggcaagta tggaacagcg 5401 tattcggatg ctgcagcaga ccatagaaca agcgttactt gaacaaggtc gcatcaccga 5461 aaaaactaac caaaactttg aatgagtcga cctgcaggca tgcaagcttt aactttaaga 5521 aggagatata cgctagcatg tttgaaccaa tggaacttac caatgacgcg gtgattaaag 5581 tcatcggcgt cggcggcggc ggcggtaatg ctgttgaaca catggtgcgc gagcgcattg 5641 aaggtgttga attcttcgcg gtaaataccg atgcacaagc gctgcgtaaa acagcggttg 5701 gacagacgat tcaaatcggt agcggtatca ccaaaggact gggcgctggc gctaatccag 5761 aagttggccg caatgcggct gatgaggatc gcgatgcatt gcgtgcggcg ctggaaggtg 5821 cagacatggt ctttattgct gcgggtatgg gtggtggtac cggtacaggt gcagcaccag 5881 tcgtcgctga agtggcaaaa gatttgggta tcctgaccgt tgctgtcgtc actaagcctt 5941 tcaactttga aggcaagaag cgtatggcat tcgcggagca ggggatcact gaactgtcca 6001 agcatgtgga ctctctgatc actatcccga acgacaaact gctgaaagtt ctgggccgcg 6061 gtatctccct gctggatgcg tttggcgcag cgaacgatgt actgaaaggc gctgtgcaag 6121 gtatcgctga actgattact cgtccgggtt tgatgaacgt ggactttgca gacgtacgca 6181 ccgtaatgtc tgagatgggc tacgcaatga tgggttctgg cgtggcgagc ggtgaagacc 6241 gtgcggaaga agctgctgaa atggctatct cttctccgct gctggaagat atcgacctgt 6301 ctggcgcgcg cggcgtgctg gttaacatca cggcgggctt cgatctgcgt ctggatgagt 6361 tcgaaacggt aggtaacacc atccgtgcat ttgcttccga caacgcgact gtggttatcg 6421 gtacttctct tgacccggat atgaatgacg agctgcgcgt aaccgttgtt gcgacaggta 6481 tcggcatgga caaacgtcct gaaatcactc tggtgaccaa taagcaggtt cagcagccag 6541 tgatggatcg ctaccagcag catgggatgg ctccgctgac ccaggagcag aagccggttg 6601 ctaaagtcgt gaatgacaat gcgccgcaaa ctgcgaaaga gccggattat ctggatatcc 6661 cagcattcct gcgtaagcaa gctgatggat ccgctggctc cgctgctggt tctggcgaat 6721 tcatggtgag caagggcgag gaggataaca tggccatcat taaggagttc atgcgcttca 6781 aggtgcacat ggaggggtcc gtgaacggcc acgtgttcga gatcgagggc gagggcgagg 6841 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc 6901 ccttcacctg ggacatcctg tcccctcaat tcatgtacgg ctccaatgcc tacgtgaagc 6961 accccgccga catccccgac tactttaagc tgtccttccc cgagggcttc aagtgggagc 7021 gcgtgatgaa attcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg 7081 acggtgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg 7141 taatgcagaa gaagaccatg ggctgggagg ccctctccga gcggatgtac cccgaggacg 7201 gcgccctgaa gggcgaggtc aagccgagag tgaagctgaa ggacggcggc cactacgacg 7261 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg 7321 tcaaccgcaa gttggacatc acctcacaca acgaggacta caccatcgtg gaacagtacg 7381 aacgtgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taa //