LOCUS BBa_K1362095 3446 bp DNA linear 06-May-2024 DEFINITION pSB4A50: Low copy BioBrick cloning/expression backbone carrying Amp resistance ACCESSION BBa_K1362095 VERSION BBa_K1362095.1 FEATURES Location/Qualifiers misc_feature 1..21 /label=Suffix misc_feature 2..7 /label=SpeI misc_feature 16..21 /label=PstI misc_feature 9..16 /label=NotI misc_feature 1..21 /label=BBa_G00001 misc_feature 22..26 /label=Nt.BstNB I recognition site misc_feature 27..29 /label=Arbitrary sequence (could be any bases) misc_feature 30..34 /label=Topoisomerase I recognition site misc_feature 30..30 /label=Generated T overhang misc_feature 22..34 /label=BBa_B0044 misc_feature 36..38 /label=stop codon (+2 frame) misc_feature 35..37 /label=stop codon (-1 frame) misc_feature 40..42 /label=stop codon (+3 frame) misc_feature 44..46 /label=stop codon (+1 frame) misc_feature 43..45 /label=stop codon (-2 frame) misc_feature 39..41 /label=stop codon (-3 frame) misc_feature 35..46 /label=BBa_B0042 stem_loop 57..88 /label=stem loop misc_feature 50..89 /label=identical to E. coli MG1655 between tonB and yciA terminator 47..115 /label=BBa_B0054 primer_bind 116..135 /label=VR misc_feature 116..135 /label=BBa_G00102 stem_loop 149..168 /label=stem_loop terminator 136..176 /label=BBa_B0062 misc_feature 177..183 /label=BBa_G0003 stem_loop 229..258 /label=Terminator misc_feature 320..687 /label=par region misc_feature 322..335 /label=inverted repeat misc_feature 341..345 /label=Tn3 integration site misc_feature 348..364 /label=inverted repeat misc_feature 375..398 /label=inverted repeat misc_feature 404..421 /label=inverted repeat misc_feature 818..900 /label=AT rich region misc_feature 899..918 /label=direct repeat misc_feature 920..939 /label=direct repeat misc_feature 952..971 /label=direct repeat misc_feature 997..1002 /label=mutated SpeI site promoter 1011..1016 /label=-35 misc_feature 1021..1042 /label=inverted repeat promoter 1029..1048 /label=promoter region promoter 1033..1038 /label=-10 misc_feature 1045..1067 /label=inverted repeat RBS 1053..1065 /label=RBS RBS 1062..1067 /label=RBS CDS 1069..2019 /label=repA misc_feature 1838..1838 /label=G to A difference with canonical pSC101 vector sequence misc_feature 1933..1933 /label=C to T difference with canonical pSC101 vector sequence misc_feature 2030..2044 /label=inverted repeat misc_feature 2095..2095 /label=G to C difference with canonical pSC101 vector sequence polyA_site 2130..2145 /label=putative polyA misc_feature 2146..2162 /label=inverted repeat misc_feature 2185..2185 /label=T to C difference with canonical pSC101 vector sequence misc_feature 184..2220 /label=BBa_I50042 misc_feature 2221..2228 /label=BBa_G0002 CDS 2308..3171 /label=ampicillin resistance promoter 2259..2264 /label=ampR -35 promoter 2285..2290 /label=ampR -10 RBS 2299..2303 /label=ampR RBS misc_feature 2229..3171 /label=BBa_P1002 misc_feature 3172..3178 /label=BBa_G0004 stem_loop 3187..3221 /label=stem_loop terminator 3179..3250 /label=BBa_B0053 primer_bind 3251..3270 /label=VF2 primer 3251..3270 /label=BBa_G00100 stem_loop 3306..3327 /label=stem loop from bacteriophage T3 misc_feature 3298..3336 /label=identical to bacteriophage T3 terminator 3271..3348 /label=BBa_B0055 misc_feature 3350..3352 /label=stop codon (+2 frame) misc_feature 3349..3351 /label=stop codon (-1 frame) misc_feature 3354..3356 /label=stop codon (+3 frame) misc_feature 3358..3360 /label=stop codon (+1 frame) misc_feature 3357..3359 /label=stop codon (-2 frame) misc_feature 3353..3355 /label=stop codon (-3 frame) misc_feature 3349..3360 /label=BBa_B0042 misc_feature 3361..3365 /label=Topoisomerase I recognition site misc_feature 3369..3373 /label=Nt.BstNB I recognition site misc_feature 3366..3368 /label=Arbitrary sequence (could be any bases) misc_feature 3365..3365 /label=Generated T overhang misc_feature 3361..3373 /label=BBa_B0043 promoter 3379..3397 /label=strong T7 promotor misc_feature 3397..3424 /label=lacO reg misc_feature 3374..3379 /label=scar of cloning into EcoRI site promoter 3374..3424 /label=BBa_K1362463 misc_feature 3425..3430 /label=EcoRI site misc_feature 3440..3445 /label=XbaI site misc_feature 3425..3446 /label=Prefix misc_feature 3431..3438 /label=NotI site misc_feature 3425..3446 /label=BBa_G00000 ORIGIN 1 tactagtagc ggccgctgca ggagtcacta agggttagtt agttagatta gcagaaagtc 61 aaaagcctcc gaccggaggc ttttgactaa aacttccctt ggggttatca ttggggctca 121 ctcaaaggcg gtaatcagat aaaaaaaatc cttagctttc gctaaggatg atttctgcta 181 gagctgtcag accaagttta cgagctcgct tggactcctg ttgatagatc cagtaatgac 241 ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg 301 tgagaatcca agcactaggg acagtaagac gggtaagcct gttgatgata ccgctgcctt 361 actgggtgca ttagccagtc tgaatgacct gtcacgggat aatccgaagt ggtcagactg 421 gaaaatcaga gggcaggaac tgctgaacag caaaaagtca gatagcacca catagcagac 481 ccgccataaa acgccctgag aagcccgtga cgggcttttc ttgtattatg ggtagtttcc 541 ttgcatgaat ccataaaagg cgcctgtagt gccatttacc cccattcact gccagagccg 601 tgagcgcagc gaactgaatg tcacgaaaaa gacagcgact caggtgcctg atggtcggag 661 acaaaaggaa tattcagcga tttgcccgag cttgcgaggg tgctacttaa gcctttaggg 721 ttttaaggtc tgttttgtag aggagcaaac agcgtttgcg acatcctttt gtaatactgc 781 ggaactgact aaagtagtga gttatacaca gggctgggat ctattctttt tatctttttt 841 tattctttct ttattctata aattataacc acttgaatat aaacaaaaaa aacacacaaa 901 ggtctagcgg aatttacaga gggtctagca gaatttacaa gttttccagc aaaggtctag 961 cagaatttac agatacccac aactcaaagg aaaaggacat gtaattatca ttgactagcc 1021 catctcaatt ggtatagtga ttaaaatcac ctagaccaat tgagatgtat gtctgaatta 1081 gttgttttca aagcaaatga actagcgatt agtcgctatg acttaacgga gcatgaaacc 1141 aagctaattt tatgctgtgt ggcactactc aaccccacga ttgaaaaccc tacaaggaaa 1201 gaacggacgg tatcgttcac ttataaccaa tacgctcaga tgatgaacat cagtagggaa 1261 aatgcttatg gtgtattagc taaagcaacc agagagctga tgacgagaac tgtggaaatc 1321 aggaatcctt tggttaaagg ctttgagatt ttccagtgga caaactatgc caagttctca 1381 agcgaaaaat tagaattagt ttttagtgaa gagatattgc cttatctttt ccagttaaaa 1441 aaattcataa aatataatct ggaacatgtt aagtcttttg aaaacaaata ctctatgagg 1501 atttatgagt ggttattaaa agaactaaca caaaagaaaa ctcacaaggc aaatatagag 1561 attagccttg atgaatttaa gttcatgtta atgcttgaaa ataactacca tgagtttaaa 1621 aggcttaacc aatgggtttt gaaaccaata agtaaagatt taaacactta cagcaatatg 1681 aaattggtgg ttgataagcg aggccgcccg actgatacgt tgattttcca agttgaacta 1741 gatagacaaa tggatctcgt aaccgaactt gagaacaacc agataaaaat gaatggtgac 1801 aaaataccaa caaccattac atcagattcc tacctacata acggactaag aaaaacacta 1861 cacgatgctt taactgcaaa aattcagctc accagttttg aggcaaaatt tttgagtgac 1921 atgcaaagta agtatgatct caatggttcg ttctcatggc tcacgcaaaa acaacgaacc 1981 acactagaga acatactggc taaatacgga aggatctgag gttcttatgg ctcttgtatc 2041 tatcagtgaa gcatcaagac taacaaacaa aagtagaaca actgttcacc gttacatatc 2101 aaagggaaaa ctgtccatat gcacagatga aaacggtgta aaaaagatag atacatcaga 2161 gcttttacga gtttttggtg cattcaaagc tgttcaccat gaacagatcg acaatgtaac 2221 tactagagct aaatacattc aaatatctat ccgctcatga gacaataacc ctgataaatg 2281 cttcaataat attgaaaaag gaagaatatg agtattcaac atttccgtgt cgcccttatt 2341 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta 2401 aaagatgccg aagatcagtt gggtgcacgt gtgggttaca tcgaactgga cctcaacagc 2461 ggtaagattc ttgagagttt tcgccccgaa gaacgtttcc caatgatgag cacttttaaa 2521 gttctgctct gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc 2581 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt 2641 acggacggca tgacagtacg cgaattatgc agcgctgcca taaccatgag tgataacacg 2701 gcggccaact tacttctgac aacgatcgga ggaccgaagg agcttaccgc ttttttgcac 2761 aacatgggtg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata 2821 ccaaacgacg agcgtgacac cacgatgcct gtagctatgg caacaacgtt gcgcaaactc 2881 ttaactggcg aacttcttac tctcgcttcc cggcaacaat taatagactg gatggaggcg 2941 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat 3001 aaatctggag ccggtgagcg tgggtcccgc ggtattattg cagccctggg gccagatggt 3061 aagccctccc gtatcgtagt tatctacacg acggggagcc aggcaactat ggacgaacgt 3121 aatcgccaga tcgctgagat aggtgcctcc ctgattaagc attggtaata atactagctc 3181 cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt 3241 acttcgcgtt tgccacctga cgtctaagaa aaggaatatt cagcaatttg cccgtgccga 3301 agaaaggccc acccgtgaag gtgagccagt gagttgattg ctacgtaatt agttagttag 3361 cccttagtga ctcgaattta atacgactca ctatagggga attgtgagcg gataacaatt 3421 ccccgaattc gcggccgctt ctagag //