LOCUS BBa_K510044 8490 bp DNA linear 22-Nov-2024 DEFINITION pUC18R6KT-miniTn7BB-Gm-bistable_switch ACCESSION BBa_K510044 VERSION BBa_K510044.1 FEATURES Location/Qualifiers misc_feature 2..21 /label=Suffix misc_feature 182..347 /label=Tn7-L end primer_bind 157..176 /label=VR (BBa_G00101) primer binding site CDS 1714..2574 /label=Ampicillin resistance (bla) misc_feature 929..1310 /label=R6K replication origin misc_feature 646..905 /label=oriT misc_feature 348..360 /label=SfiI restriction site misc_feature 3176..3374 /label=Tn7-R end misc_feature 3385..3479 /label=double terminator (BBa_B0014) misc_feature 3490..3537 /label=Flipase recombination sequence (FRT) misc_feature 3163..3175 /label=SfiI restriction site misc_feature 3548..3553 /label=NcoI restriction site CDS 3554..4087 /label=Gentamicin resistance misc_feature 3542..3547 /label=SphI restriction site misc_feature 4345..4350 /label=NcoI restriction site misc_feature 4351..4355 /label=SphI restriction site misc_feature 4528..4549 /label=Prefix primer_bind 4410..4429 /label=VF2 (G00100) primer binding site misc_feature 4357..4404 /label=Flipase recombination sequence (FRT) stem_loop 22..93 /label=E. coli his operon terminator stem_loop 4487..4520 /label=Terminator misc_feature 1..4549 /label=BBa_K510012 CDS 4558..4645 /label=end of LacI coding region (inactive) misc_feature 4558..4757 /label=BBa_R0010 misc_feature 4646..4683 /label=CAP binding site promoter 4694..4699 /label=-35 promoter 4718..4723 /label=-10 misc_feature 4723..4757 /label=LacI binding site misc_feature 4730..4730 /label=start promoter 4558..4757 /label=BBa_R0010 RBS 4766..4778 /label=RBS-3\Weak RBS 4772..4775 /label=RBS misc_feature 4766..4778 /label=BBa_B0032 RBS 4766..4778 /label=BBa_B0032 CDS 4785..5498 /label=cIts CDS 4785..5498 /label=BBa_K177050 RBS 5507..5521 /label=RBS-1\Strong RBS 5514..5518 /label=RBS misc_feature 5507..5521 /label=BBa_B0030 RBS 5507..5521 /label=BBa_B0030 CDS 5528..6247 /label=GFP protein CDS 5528..6247 /label=BBa_E0040 stem_loop 6267..6310 /label=stem_loop misc_feature 6256..6335 /label=BBa_B0010 terminator 6256..6335 /label=BBa_B0010 stem_loop 6351..6370 /label=T7 TE misc_feature 6377..6377 /label=stop polyA_site 6371..6384 /label=polya misc_feature 6344..6384 /label=BBa_B0012 terminator 6344..6384 /label=BBa_B0012 misc_feature 4558..6384 /label=BBa_K177034 promoter 6430..6435 /label=-10 promoter 6407..6412 /label=-35 misc_feature 6417..6433 /label=OR1 misc_feature 6393..6409 /label=OR2 misc_feature 6393..6441 /label=BBa_R0051 promoter 6393..6441 /label=BBa_R0051 RBS 6450..6462 /label=RBS-3\Weak RBS 6456..6459 /label=RBS misc_feature 6450..6462 /label=BBa_B0032 RBS 6450..6462 /label=BBa_B0032 misc_feature 7558..7596 /label=LVA CDS 6469..7596 /label=lacI-LVA misc_feature 6469..7596 /label=BBa_C0012 misc_feature 7597..7621 /label=Help:Barcodes CDS 6469..7596 /label=BBa_C0012 misc_feature 6393..7621 /label=BBa_K177012 misc_feature 7634..7637 /label=conserved RBS 7630..7641 /label=BBa_B0034 CDS 7648..8322 /label=mrfp1 misc_feature 8329..8353 /label=Help:Barcodes CDS 7648..8353 /label=BBa_E1010 stem_loop 8373..8416 /label=stem_loop misc_feature 8362..8441 /label=BBa_B0010 terminator 8362..8441 /label=BBa_B0010 stem_loop 8457..8476 /label=T7 TE misc_feature 8483..8483 /label=stop polyA_site 8477..8490 /label=polya misc_feature 8450..8490 /label=BBa_B0012 terminator 8450..8490 /label=BBa_B0012 misc_feature 7630..8490 /label=BBa_K094130 misc_feature 4558..8490 /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 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 481 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgttaag gtatactttc 661 cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt 721 cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg 781 gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac 841 tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccgat 901 aagctctgat aacgcaggaa agaacatggg cagttcaacc tgttgatagt acgtactaag 961 ctctcatgtt tcacgtacta agctctcaag tttaacgtac taagctctca tgtttaacga 1021 actaaaccct catggctaac gtactaagct ctcatggcta acgtactaag ctctcatgtt 1081 tcacgtacta agctctcatg tttgaacaat aaaattaata taaatcagca acttaaatag 1141 cctctaaggt tttaagtttt ataagaaaaa aaagaatata taaggctttt aaagctttta 1201 aggtttaacg gttgtggaca acaagccagg gatgtaacgc actgagaagc ccttagagcc 1261 tctcaaagca attttgagtg acacaggaac acttaacggc tgacaaccgg ctggtaacag 1321 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 1381 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 1441 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 1501 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 1561 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 1621 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 1681 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 1741 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 1801 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 1861 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 1921 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 1981 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 2041 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 2101 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 2161 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 2221 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 2281 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 2341 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 2401 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 2461 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 2521 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 2581 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 2641 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 2701 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 2761 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 2821 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 2881 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 2941 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 3001 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 3061 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 3121 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccgcct aggcctgtgg 3181 gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa 3241 actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg 3301 acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt 3361 aaagaggggc gtggggttcg cttgtcacac tggctcacct tcgggtgggc ctttctgcgt 3421 ttatatacta gagagagaat ataaaaagcc agattattaa tccggctttt ttattatttg 3481 ctagatcttg aagtacctat tccgaagttc ctattctcta aaaagtatag gaacttcaga 3541 ggcatgccca tggttaggtg gcggtacttg ggtcgatatc aaagtgcatc acttcttccc 3601 gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct gcttgcacgt 3661 agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg agcgcggtgg 3721 caatgccctg cctccggtgc tcgccggaga ctgcgagatc atagatatag atctcactac 3781 gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc gcttcttggt 3841 cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg taatcggagt 3901 ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa agatcaagag 3961 cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg atgcccatac 4021 ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc 4081 gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc 4141 ttggatgccc gaggcataga ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc 4201 actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata 4261 cgctacttgc attacagttt acgaaccgaa caggcttatg tcaattcgat ccattgctgt 4321 tgacaaaggg aatcagggga tcttccatgg gcatgcgaag tacctattcc gaagttccta 4381 ttctctaaaa agtataggaa cttcagagct gccacctgac gtctaagaaa ccattattat 4441 catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat aaaaaaaatc 4501 cttagctttc gctaaggatg atttctggaa ttcgcggccg cttctagagt actagagcaa 4561 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt 4621 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt 4681 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg 4741 gataacaatt tcacacatac tagagtcaca caggaaagta ctagatgagc acaaaaaaga 4801 aaccattaac acaagagcag cttgaggacg cacgtcgcct taaagcaatt tatgaaaaaa 4861 agaaaaatga acttggctta tcccaggaat ctgtcgcaga caagatgggg atggggcagt 4921 caggcgttgg tgctttattt aatggcatca atgcattaaa tgcttataac gccgcattgc 4981 ttacaaaaat tctcaaagtt agcgttgaag aatttagccc ttcaatcgcc agagaaatct 5041 acgagatgta tgaagcggtt agtatgcagc cgtcacttag aagtgagtat gagtaccctg 5101 ttttttctca tgttcaggca gggatgttct cacctgagct tagaaccttt accaaaggtg 5161 atgcggagag atgggtaagc acaaccaaaa aagccagtga ttctgcattc tggcttgagg 5221 ttgaaggtaa ttccatgacc gcaccaacag gctccaagcc aagctttcct gacggaatgt 5281 taattctcgt tgaccctgag caggctgttg agccaggtga tttctgcata gccagacttg 5341 ggggtgatga gtttaccttc aagaaactga tcagggatag cggtcaggtg tttttacaac 5401 cactaaaccc acagtaccca atgatcccat gcaatgagag ttgttccgtt gtggggaaag 5461 ttatcgctag tcagtggcct gaagagacgt ttggctgata ctagagatta aagaggagaa 5521 atactagatg cgtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt 5581 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac 5641 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc 5701 aacacttgtc actactttcg gttatggtgt tcaatgcttt gcgagatacc cagatcatat 5761 gaaacagcat gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat 5821 atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac 5881 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg 5941 acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa 6001 gaatggaatc aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact 6061 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa 6121 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat 6181 ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa 6241 ataataatac tagagccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct 6301 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt 6361 cgggtgggcc tttctgcgtt tatatactag agtaacaccg tgcgtgttga ctattttacc 6421 tctggcggtg ataatggttg ctactagagt cacacaggaa agtactagat ggtgaatgtg 6481 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc 6541 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg 6601 atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg 6661 ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg 6721 gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga 6781 agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg 6841 ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact 6901 aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc 6961 tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa 7021 atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg 7081 cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt 7141 gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg 7201 atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg 7261 ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt 7321 tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg 7381 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc 7441 tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg 7501 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcaggct 7561 gcaaacgacg aaaactacgc tttagtagct taataactct gatagtgcta gtgtagatct 7621 ctactagaga aagaggagaa atactagatg gcttcctccg aagacgttat caaagagttc 7681 atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt 7741 gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt 7801 ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct 7861 tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc 7921 aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc 7981 tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc 8041 gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac 8101 ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt 8161 cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt 8221 gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt 8281 gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaataacg ctgatagtgc 8341 tagtgtagat cgctactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact 8401 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct 8461 caccttcggg tgggcctttc tgcgtttata //