LOCUS Exported 3488 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Start-Stop Assembly Level 1DE plasmid. ACCESSION . VERSION . KEYWORDS pStA1DE 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" promoter 1..31 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 39..55 /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 44..66 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 63..79 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 63..79 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 75..398 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" misc_feature 92..148 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(149..166) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(149..165) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(158..180) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(374..393) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" terminator 426..512 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(915..932) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1049..1594) /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(1129..1148) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 1706..1734 /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" CDS 1782..2972 /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(2134..2153) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" terminator 3310..3404 /label=lambda t0 terminator /note="transcription terminator from phage lambda" protein_bind 3453..3474 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 61 cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag 121 aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg 181 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc 241 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg 301 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat 361 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt tccgctcttc aggacgctag 421 agaccatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc 481 cagtctttcg actgagcctt tcgttttatt tgatgcccga aagggcctcg tgatacgcca 541 aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa 601 gtgatcttcc gtcacaggta tttattcggc gcaaagtgtg tcgggtgatg ctgccaactt 661 actgatttag tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt tctatcagct 721 gtccctcctg ttcagctact gacggggtgg tgcgtaacgg caaaagcacc gccggacatc 781 agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt 841 gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg 901 atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcgacgagcg 961 gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg 1021 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc 1081 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg 1141 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg 1201 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag 1261 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc 1321 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca 1381 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta 1441 aactgaaagg acaagttttg gtgactgcgc tcctccaagc catttacctc ggttcaaaga 1501 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga 1561 gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa 1621 atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat 1681 acgatataag ttgtaattct catgtttgac agcttatcat cgataagctt taatgcggta 1741 gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc 1801 tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg 1861 tactgccggg cctcttgcgg gatatcgtcc attccgacag catcgccagt cactatggcg 1921 tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc acccgttctc ggagcactgt 1981 ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct acttggagcc actatcgact 2041 acgcgatcat ggcgaccaca cccgtcctgt ggatcctcta cgccggacgc atcgtggccg 2101 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg 2161 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag 2221 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg 2281 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg 2341 gagagcgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc 2401 ggggcatgac tatcgtcgcc gcacttatga ctgttttctt tatcatgcaa ctcgtaggac 2461 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga 2521 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca 2581 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg 2641 acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta 2701 tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc 2761 aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa 2821 cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga 2881 acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc 2941 gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg 3001 attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac 3061 caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat 3121 ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac 3181 ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg 3241 aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcgccgcagc cggacgaccg 3301 agcgccttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt 3361 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatggtctc agcttcagag 3421 aagagccagt agggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca 3481 ccccaggc //