LOCUS Exported 2658 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Start-Stop Assembly Level 0 plasmid. ACCESSION . VERSION . KEYWORDS pStA0 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2658) AUTHORS Taylor GM, Mordaka PM, Heap JT TITLE Start-Stop Assembly: a functionally scarless DNA assembly system optimized for metabolic engineering. JOURNAL Nucleic Acids Res. 2018 Nov 20. pii: 5193345. doi: 10.1093/nar/gky1182. PUBMED 30462270 REFERENCE 2 (bases 1 to 2658) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2658 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 1..95 /label=lambda t0 terminator /note="transcription terminator from phage lambda" protein_bind 134..155 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 170..200 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 208..224 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="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 213..235 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 232..248 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 232..248 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 244..567 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" misc_feature 261..317 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(318..335) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(318..334) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(327..349) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(543..562) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" terminator 592..678 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(691..709) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 777..881 /gene="bla" /label=AmpR promoter CDS 882..1742 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1100..1119) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1913..2501 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2402..2421 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc 61 tcggttgccg ccgggcgttt tttattggtg agaatccagt gtgagagacc aagggcagtg 121 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta 181 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca 241 gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga ggatccccgg 301 gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc 361 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa 421 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg 481 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc 541 agtacaatct gctctgatgc cgcatagggt ctcagacctc tagggcggcg gatttgtcct 601 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg 661 agcctttcgt tttatttgat gcccgaaagg gcctcgtgat acgcctattt ttataggtta 721 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg 781 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 841 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 901 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 961 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac 1021 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga 1081 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 1141 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca 1201 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 1261 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa 1321 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 1381 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 1441 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag 1501 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 1561 ggtttattgc tgataaatct ggagccggtg agcgtgggtc ccgcggtatc attgcagcac 1621 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 1681 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 1741 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat 1801 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 1861 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 1921 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 1981 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 2041 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 2101 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 2161 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 2221 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 2281 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 2341 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 2401 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 2461 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 2521 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 2581 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc 2641 gaacgaccga gcgccttg //