LOCUS Exported 3488 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Start-Stop Assembly Level 1EZ plasmid. ACCESSION . VERSION . KEYWORDS pStA1EZ SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3488) 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 3488) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3488 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 1..95 /label=lambda t0 terminator /note="transcription terminator from phage lambda" protein_bind 144..165 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 180..210 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 218..234 /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 223..245 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 242..258 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 242..258 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 254..577 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" misc_feature 271..327 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(328..345) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(328..344) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(337..359) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(553..572) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" terminator 605..691 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(1094..1111) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1228..1773) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(1308..1327) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 1885..1913 /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" CDS 1961..3151 /codon_start=1 /gene="tet" /product="tetracycline efflux protein" /label=TcR /note="confers resistance to tetracycline" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST" primer_bind complement(2313..2332) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc 61 tcggttgccg ccgggcgttt tttattggtg agaatggtct cacgctcaga gaagagccag 121 tagggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct 181 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac 241 acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga 301 ggatccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga 361 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg 421 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga 481 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg 541 gtgcactctc agtacaatct gctctgatgc cgcatagttt ccgctcttca ggatactaga 601 gaccatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc 661 agtctttcga ctgagccttt cgttttattt gatgcccgaa agggcctcgt gatacgccaa 721 aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag 781 tgatcttccg tcacaggtat ttattcggcg caaagtgtgt cgggtgatgc tgccaactta 841 ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt ctatcagctg 901 tccctcctgt tcagctactg acggggtggt gcgtaacggc aaaagcaccg ccggacatca 961 gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt cagtgaagtg 1021 cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga 1081 tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg 1141 aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg 1201 aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca 1261 cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc 1321 gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt 1381 cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg 1441 cagttsgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct 1501 tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag 1561 cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa 1621 actgaaagga caagttttgg tgactgcgct cctccaagcc atttacctcg gttcaaagag 1681 ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag 1741 caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa 1801 tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata 1861 cgatataagt tgtaattctc atgtttgaca gcttatcatc gataagcttt aatgcggtag 1921 tttatcacag ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct 1981 catcgtcatc ctcggcaccg tcaccctgga tgctgtaggc ataggcttgg ttatgccggt 2041 actgccgggc ctcttgcggg atatcgtcca ttccgacagc atcgccagtc actatggcgt 2101 gctgctagcg ctatatgcgt tgatgcaatt tctatgcgca cccgttctcg gagcactgtc 2161 cgaccgcttt ggccgccgcc cagtcctgct cgcttcgcta cttggagcca ctatcgacta 2221 cgcgatcatg gcgaccacac ccgtcctgtg gatcctctac gccggacgca tcgtggccgg 2281 catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga 2341 agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg 2401 ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc 2461 ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg 2521 agagcgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg 2581 gggcatgact atcgtcgccg cacttatgac tgttttcttt atcatgcaac tcgtaggaca 2641 ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat 2701 gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac 2761 tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggccga 2821 cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat 2881 gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca 2941 ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta ccagcctaac 3001 ttcgatcatt ggaccgctga tcgtcacggc gatttatgcc gcctcggcga gcacatggaa 3061 cgggttggca tggattgtag gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg 3121 cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct cgctaacgga 3181 ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc 3241 aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc 3301 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc 3361 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga 3421 acgtgaagcg actgctgctg caaaacgtct gcgacctgag cgccgcagcc ggacgaccga 3481 gcgccttg //