LOCUS pSB2K4 4467 bp DNA linear 15-May-2024 DEFINITION pSB2K3-derived BioBrick plasmid free of many restriction sites ACCESSION pSB2K4 VERSION pSB2K4.1 FEATURES Location/Qualifiers misc_feature 1..21 /label=BioBrick suffix stem_loop 51..120 /label=E. coli his operon terminator stem_loop 59..91 /label=Stem loop misc_feature 93..93 /label=T->C misc_feature 146..146 /label=C->A primer_bind 184..203 /label=Verification reverse misc_feature 269..317 /label=f1 ori misc_feature 292..292 /label=T->A misc_feature 394..412 /label=repE binding site misc_feature 462..462 /label=A->G mutation misc_feature 477..495 /label=repE binding site misc_feature 394..641 /label=incC misc_feature 551..551 /label=T->A mutation misc_feature 568..586 /label=repE binding site misc_feature 578..578 /label=T->C misc_feature 597..619 /label=repE binding site misc_feature 623..641 /label=repE binding site misc_feature 692..692 /label=A->C misc_feature 857..857 /label=G->C CDS 644..1399 /label=repE misc_feature 1091..1091 /label=G->C misc_feature 1100..1100 /label=T->C misc_feature 1142..1142 /label=C->T misc_feature 1426..1426 /label=Predicted transcription start promoter 1426..1431 /label=Possible -35 promoter 1433..1441 /label=Possible -10 promoter 1444..1452 /label=Possible -10 misc_feature 1447..1447 /label=Predicted transcription start misc_feature 1454..1454 /label=G->T promoter 1455..1463 /label=Possible -10 promoter 1455..1460 /label=Possible -35 misc_feature 1460..1460 /label=Predicted transcription start promoter 1477..1482 /label=Possible -35 misc_feature 1497..1515 /label=repE binding site misc_feature 1527..1537 /label=repE binding site misc_feature 1552..1570 /label=repE binding site misc_feature 1581..1591 /label=repE binding site misc_feature 1490..1706 /label=oriS misc_feature 1636..1682 /label=Low GC region misc_feature 1823..1844 /label=Low GC region promoter 2064..2238 /label=lacOP misc_feature 2565..2565 /label=Maybe A or G misc_feature 2567..2567 /label=T->G misc_feature 2699..2699 /label=T->G CDS 2316..3161 /label=repL misc_feature 2774..2774 /label=T->C misc_feature 3017..3017 /label=G->A misc_feature 3251..3251 /label=T->C promoter 3269..3314 /label=nnpp promoter 3 (0.95)' promoter 3359..3364 /label=Predicted -35 promoter 3353..3398 /label=nnpp promoter 2 (0.96)' promoter 3379..3387 /label=Predicted -10 misc_feature 3394..3394 /label=Predicted transcription start promoter 3375..3420 /label=nnpp promoter 1 (.92)' misc_feature 3507..3507 /label=T->A misc_feature 3702..3702 /label=T->C misc_feature 3849..3849 /label=G->C CDS 3460..4275 /label=kanR misc_feature 3903..3903 /label=T->G misc_feature 4119..4119 /label=C->T primer_bind 4313..4332 /label=VF2 (not an exact match, but works) misc_feature 4321..4321 /label=G->C stem_loop 4389..4392 /label=end of terminator stem_loop 4402..4421 /label=Stem loop misc_feature 4446..4467 /label=BioBrick prefix ORIGIN 1 tactagtagc ggccgctgca gcaccggtgg cagttcgaat gatcatgcag tccggcaaaa 61 aagggcaagg tgtcaccacc ctgccctttt tccttaaaac cgaaaagatt acttcgcgtt 121 atgcaggctt cctcgctcac tgactagctg cgctcggtcg ttcggctgcg gcgagcggta 181 tcagctcact caaaggcggt aatggcgcgg ctcgcgtacc cataataccc ataatagctg 241 tttgccatcg cgtatgcatc gatcacgtgt ccacgttctt taatagtgga cacttgttcc 301 aaactggaac aacactcgga tcgaaccgcc gcgcaccgtg ggaaaaactc caggtagagg 361 tacacacgcg gatagccaat tcagagtaat aaactgtgat aatcaaccct catcaatgat 421 gacgaactaa cccccgatat caggtcacat gacgaaggga aggagaagga aatcaactgt 481 gacaaactgc cctcaaattt ggcttcctta aaaattacag ttcaaaaagt atgagaaaat 541 ccatgcaggc agaaggaaac agcaaaactg tgacaaacta ccctcagtag gtcagaacaa 601 atgtgacgaa ccaccctcaa atctgtgaca gataaccctc agactatcct gtcgtcatgg 661 aagtgatatc gcggaaggaa aatacgatat gcgtcgtctg gcggcctttc tttttctcaa 721 tgtatgagag gcgcattgga gttctgctgt tgatctcatt aacacagact tgcaggaagc 781 ggcggcggaa gtcaggcata cgctggtaac tttgaggcag ctggtaacgc tctatgatcc 841 agtcgatttt cagagacacg atgcctgagc catccggctt acgatactga cacagggatt 901 cgtataaacg catggcatac ggattggtga tttcttttgt ttcactaagc cgaaactgcg 961 taaaccgatt ctgtaacccg ataaagaagg gaatgagata tgggttgata tgtacactgt 1021 aaagccctct ggatggactg tgcgcacgtt tgataaacca aggaaaagat tcatagcctt 1081 tttcatcacc cgcatcctcc tcagggcgat aaaaaaccac ttccttcccc gcgaaactct 1141 ttaatgcctg ccgtatatcc ttactggctt ccgcagaggt caatccgaat atttcagcat 1201 atttagcaac atggatctcg cagataccgt catgttcctg tagggtgcca tcagattttc 1261 tgatctggtc aacgaacaga tacagcatac gtttttgatc ccgagagaga ctatatgccg 1321 cctcagtgag gtcgtttgac tggacgattc gcgggctatt tttacgtttc ttgtgattga 1381 taaccgctgt ttccgccatg acagatccat gtgaagtgtg acaagttttt agattgtcac 1441 actaaataaa aaatagtcaa taagcaggga taactttgtg aaaaaacagc ttcttctgag 1501 ggcaatttgt cacagggtta agggcaattt gtcacagaca ggactgtcat ttgagggtga 1561 tttgtcacac tgaaagggca atttgtcaca acaccttctg tagaaccagc atggataaag 1621 gcctacaagg cgctctaaaa aagaagatct aaaaactata aaaaaaataa ttataaaaat 1681 atccccgtgg ataagtggat aaccccaagg gaagtttttt caggcatcgt gtgtaagcag 1741 aatatataag tgctgttccc tggtgcttcc tcgctcactc gaaattgccg gggatagctt 1801 tatgcttgta aaccgttttg tgaaaaaatt tttaaaataa aaaaggggac ctctagggtc 1861 cccaattaat tagtaatata atctattaaa ggtcattcaa aaggtcatcc accggatcaa 1921 ttcccctgct cgcgcaggct gggtgccaag ctctcgggta acatcaaggc ccgatccttg 1981 gagcccttgc cctcccgcac gatgatcgtg ccgtgatcga aatccagatc cttgacccgc 2041 agttgcaaac cctcactgat ccgattcatt aatgcagctg gcacgacagg tttcccgact 2101 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc 2161 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat 2221 ttcacacagg aaacagctat gaccatgatt acgccaagcg cggtacctgt cgcggcaacg 2281 cgctaacaga cgtagtaaga accaccagca ttgtaatgct ggctaaagtc actttcctga 2341 gctgtataac gatgagcgat tttacttttt ctggctatga attggcctgc tttgtaacac 2401 actccggtct atcccgtagc gccgggcata tcctgtcgca atgtgcaaat ctcgcggcaa 2461 caaccagtga atacttcatt cacaagcctc accgcctgat cgcggcagaa actggttata 2521 gccaatcaac cgtcgttcgt gcattccgtg aagctgtaaa caaagggatc ctgtctgtag 2581 agattgttat cggcgatcac cgtgaacgtc gcgctaacct gtatcggttt acaccatcct 2641 ttttggcctt cgcacaacaa gccaaaaatg cgctgataga aagcaaatta aagatctcgt 2701 cagcggcaac caaggttaaa gctgttctcg ctaagacatt ggctttattt aattttttat 2761 ccacaccccc atgccaaaat gataccccct ccccctgtca ggatgacgtg gcaataaaga 2821 ataagaagtc acaagttaaa aaaacaaaaa gatcagtttc cggcggtgcc ggaacaacca 2881 gcctcaaaaa attgacttca tggatcgcta aggcaaaagc aaaggctgac aatctgcggt 2941 tatccaaaaa acgcactcaa aaacatgagt tcaagcagaa agtagaggcg gctgcgcgga 3001 aatatgctta cctgaaaaac aagcgttcgc ctgatattgg cgggatatca aacttcgata 3061 acctaccgca ttgcatgacg gtaaacgaag ctcttaatgc ggttttagcc aaaaataaag 3121 ataacgaaca atggggtata ccggcaggat tcagagggta atgaattgct ctaattataa 3181 ccatgcatac tttcaacacc tctagtttgc catgaggcaa actcataggt gtcctggtaa 3241 gaggacactg ctgccaaaac tggacgcccc attattgcaa ttaataaaca actaacggac 3301 aattctacct aacaataagt ggagttgcgg cccgccgcga tccgctagga aagccacgtt 3361 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata 3421 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa 3481 acgtcttgct caaggccgcg attaaaatcc aacatggatg ctgatttata tgggtataaa 3541 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct accgattgta tgggaagccc 3601 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat 3661 gagatggtca gactaaactg gctgacggaa tttatgcctc tcccgaccat caagcatttt 3721 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc 3781 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc 3841 ctgcgccgct tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt 3901 cggctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat 3961 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa actgttgcca 4021 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac 4081 gaggggaaat taataggttg tattgatgtt ggacgagttg gaatcgcaga ccgataccag 4141 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt 4201 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc 4261 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcgagc tgtgccacct 4321 cacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 4381 cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg gtattctgcg 4441 gaccggaatt cgcggccgct tctagag //