LOCUS BBa_K510043 8329 bp DNA linear 18-Feb-2025 DEFINITION pUC18Sfi-miniTn7BB-Gm-bistable_switch ACCESSION BBa_K510043 VERSION BBa_K510043.1 FEATURES Location/Qualifiers misc_feature 2..21 /label=Suffix stem_loop 22..93 /label=E. coli his operon terminator primer_bind 157..176 /label=VR (BBa_G00101) primer binding site misc_feature 182..347 /label=Tn7-L end misc_feature 348..360 /label=SfiI restriction site CDS 966..1826 /label=Ampicillin resistance (bla) misc_feature 1965..2644 /label=ColEI (pUC18) Replication origin misc_feature 3002..3014 /label=SfiI restriction site misc_feature 3014..3213 /label=Tn7-R end misc_feature 3329..3376 /label=Flipase recombination sequence (FRT) CDS 3393..3926 /label=Gentamycin resistance misc_feature 4196..4243 /label=Flipase recombination sequence (FRT) misc_feature 4367..4387 /label=Prefix misc_feature 3224..3318 /label=double terminator (BBa_B0014) misc_feature 3381..3386 /label=SphI restriction site misc_feature 3387..3392 /label=NcoI restrictino site misc_feature 4184..4189 /label=NcoI restriction site misc_feature 4190..4195 /label=SphI restriction site primer_bind 4249..4268 /label=VF2 (G00100) primer binding site stem_loop 4326..4359 /label=Terminator misc_feature 1..4388 /label=BBa_K510000 CDS 4397..4484 /label=end of LacI coding region (inactive) misc_feature 4397..4596 /label=BBa_R0010 misc_feature 4485..4522 /label=CAP binding site promoter 4533..4538 /label=-35 promoter 4557..4562 /label=-10 misc_feature 4562..4596 /label=LacI binding site misc_feature 4569..4569 /label=start promoter 4397..4596 /label=BBa_R0010 RBS 4605..4617 /label=RBS-3\Weak RBS 4611..4614 /label=RBS misc_feature 4605..4617 /label=BBa_B0032 RBS 4605..4617 /label=BBa_B0032 CDS 4624..5337 /label=cIts CDS 4624..5337 /label=BBa_K177050 RBS 5346..5360 /label=RBS-1\Strong RBS 5353..5357 /label=RBS misc_feature 5346..5360 /label=BBa_B0030 RBS 5346..5360 /label=BBa_B0030 CDS 5367..6086 /label=GFP protein CDS 5367..6086 /label=BBa_E0040 stem_loop 6106..6149 /label=stem_loop misc_feature 6095..6174 /label=BBa_B0010 terminator 6095..6174 /label=BBa_B0010 stem_loop 6190..6209 /label=T7 TE misc_feature 6216..6216 /label=stop polyA_site 6210..6223 /label=polya misc_feature 6183..6223 /label=BBa_B0012 terminator 6183..6223 /label=BBa_B0012 misc_feature 4397..6223 /label=BBa_K177034 promoter 6269..6274 /label=-10 promoter 6246..6251 /label=-35 misc_feature 6256..6272 /label=OR1 misc_feature 6232..6248 /label=OR2 misc_feature 6232..6280 /label=BBa_R0051 promoter 6232..6280 /label=BBa_R0051 RBS 6289..6301 /label=RBS-3\Weak RBS 6295..6298 /label=RBS misc_feature 6289..6301 /label=BBa_B0032 RBS 6289..6301 /label=BBa_B0032 misc_feature 7397..7435 /label=LVA CDS 6308..7435 /label=lacI-LVA misc_feature 6308..7435 /label=BBa_C0012 misc_feature 7436..7460 /label=Help:Barcodes CDS 6308..7435 /label=BBa_C0012 misc_feature 6232..7460 /label=BBa_K177012 misc_feature 7473..7476 /label=conserved RBS 7469..7480 /label=BBa_B0034 CDS 7487..8161 /label=mrfp1 misc_feature 8168..8192 /label=Help:Barcodes CDS 7487..8192 /label=BBa_E1010 stem_loop 8212..8255 /label=stem_loop misc_feature 8201..8280 /label=BBa_B0010 terminator 8201..8280 /label=BBa_B0010 stem_loop 8296..8315 /label=T7 TE misc_feature 8322..8322 /label=stop polyA_site 8316..8329 /label=polya misc_feature 8289..8329 /label=BBa_B0012 terminator 8289..8329 /label=BBa_B0012 misc_feature 7469..8329 /label=BBa_K094130 misc_feature 4397..8329 /label=BBa_K177038 ORIGIN 1 tactagtagc ggccgctgca gtccggcaaa aaaacgggca aggtgtcacc accctgccct 61 ttttctttaa aaccgaaaag attacttcgc gttatgcagg cttcctcgct cactgactcg 121 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaataggc 181 caaccagata agtgaaatct agttccaaac tattttgtca tttttaattt tcgtattagc 241 ttacgacgct acacccagtt cccatctatt ttgtcactct tccctaaata atccttaaaa 301 actccatttc cacccctccc agttcccaac tattttgtcc gcccacaggc cgcctaggcc 361 gtagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc 421 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc 481 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt 541 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa 601 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc 661 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga 721 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg 781 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg 841 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 901 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 961 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 1021 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 1081 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 1141 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 1201 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 1261 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 1321 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 1381 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 1441 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 1501 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc 1561 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc 1621 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 1681 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 1741 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 1801 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga 1861 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 1921 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 1981 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 2041 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 2101 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 2161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 2221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 2281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 2341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagctt tgagaaagcg 2401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 2461 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 2521 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 2581 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 2641 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 2701 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag 2761 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca 2821 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga 2881 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt 2941 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgaat 3001 tggccgccta ggcctgtggg cggacaataa agtcttaaac tgaacaaaat agatctaaac 3061 tatgacaata aagtcttaaa ctagacagaa tagttgtaaa ctgaaatcag tccagttatg 3121 ctgtgaaaaa gcatactgga cttttgttat ggctaaagca aactcttcat tttctgaagt 3181 gcaaattgcc cgtcgtatta aagaggggcg tggggttcgc ttgtcacact ggctcacctt 3241 cgggtgggcc tttctgcgtt tatatactag agagagaata taaaaagcca gattattaat 3301 ccggcttttt tattatttgc tagatcttga agtacctatt ccgaagttcc tattctctaa 3361 aaagtatagg aacttcagag gcatgcccat ggttaggtgg cggtacttgg gtcgatatca 3421 aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca 3481 ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg 3541 agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca 3601 tagatataga tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg 3661 ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag 3721 ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca 3781 cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat 3841 gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc 3901 gtaacatcgt tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac 3961 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata 4021 acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga 4081 ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt 4141 caattcgatc cattgctgtt gacaaaggga atcaggggat cttccatggg catgcgaagt 4201 acctattccg aagttcctat tctctaaaaa gtataggaac ttcagagctg ccacctgacg 4261 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga 4321 atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc 4381 ttctagagta ctagagcaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa 4441 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat 4501 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg 4561 ttgtgtggaa ttgtgagcgg ataacaattt cacacatact agagtcacac aggaaagtac 4621 tagatgagca caaaaaagaa accattaaca caagagcagc ttgaggacgc acgtcgcctt 4681 aaagcaattt atgaaaaaaa gaaaaatgaa cttggcttat cccaggaatc tgtcgcagac 4741 aagatgggga tggggcagtc aggcgttggt gctttattta atggcatcaa tgcattaaat 4801 gcttataacg ccgcattgct tacaaaaatt ctcaaagtta gcgttgaaga atttagccct 4861 tcaatcgcca gagaaatcta cgagatgtat gaagcggtta gtatgcagcc gtcacttaga 4921 agtgagtatg agtaccctgt tttttctcat gttcaggcag ggatgttctc acctgagctt 4981 agaaccttta ccaaaggtga tgcggagaga tgggtaagca caaccaaaaa agccagtgat 5041 tctgcattct ggcttgaggt tgaaggtaat tccatgaccg caccaacagg ctccaagcca 5101 agctttcctg acggaatgtt aattctcgtt gaccctgagc aggctgttga gccaggtgat 5161 ttctgcatag ccagacttgg gggtgatgag tttaccttca agaaactgat cagggatagc 5221 ggtcaggtgt ttttacaacc actaaaccca cagtacccaa tgatcccatg caatgagagt 5281 tgttccgttg tggggaaagt tatcgctagt cagtggcctg aagagacgtt tggctgatac 5341 tagagattaa agaggagaaa tactagatgc gtaaaggaga agaacttttc actggagttg 5401 tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag 5461 agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa 5521 aactacctgt tccatggcca acacttgtca ctactttcgg ttatggtgtt caatgctttg 5581 cgagataccc agatcatatg aaacagcatg actttttcaa gagtgccatg cccgaaggtt 5641 atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca cgtgctgaag 5701 tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag 5761 aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac aatgtataca 5821 tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga cacaacattg 5881 aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc 5941 ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca 6001 acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg 6061 gcatggatga actatacaaa taataatact agagccaggc atcaaataaa acgaaaggct 6121 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag 6181 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atatactaga gtaacaccgt 6241 gcgtgttgac tattttacct ctggcggtga taatggttgc tactagagtc acacaggaaa 6301 gtactagatg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt 6361 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg 6421 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca 6481 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc 6541 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt 6601 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct 6661 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc 6721 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc 6781 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt 6841 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg 6901 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga 6961 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga 7021 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg 7081 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga 7141 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct 7201 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg 7261 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca 7321 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg 7381 actggaaagc gggcaggctg caaacgacga aaactacgct ttagtagctt aataactctg 7441 atagtgctag tgtagatctc tactagagaa agaggagaaa tactagatgg cttcctccga 7501 agacgttatc aaagagttca tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca 7561 cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa 7621 actgaaagtt accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt 7681 ccagtacggt tccaaagctt acgttaaaca cccggctgac atcccggact acctgaaact 7741 gtccttcccg gaaggtttca aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt 7801 taccgttacc caggactcct ccctgcaaga cggtgagttc atctacaaag ttaaactgcg 7861 tggtaccaac ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg gttgggaagc 7921 ttccaccgaa cgtatgtacc cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct 7981 gaaactgaaa gacggtggtc actacgacgc tgaagttaaa accacctaca tggctaaaaa 8041 accggttcag ctgccgggtg cttacaaaac cgacatcaaa ctggacatca cctcccacaa 8101 cgaagactac accatcgttg aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc 8161 ttaataacgc tgatagtgct agtgtagatc gctactagag ccaggcatca aataaaacga 8221 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc 8281 tactagagtc acactggctc accttcgggt gggcctttct gcgtttata //