LOCUS Exported 3233 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pUC-UAS1B4-Leum SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3233) AUTHORS Blazeck J, Liu L, Redden H, Alper H TITLE Tuning gene expression in Yarrowia lipolytica by a hybrid promoter approach. JOURNAL Appl Environ Microbiol. 2011 Nov;77(22):7905-14. doi: 10.1128/AEM.05763-11. Epub 2011 Sep 16. PUBMED 21926196 REFERENCE 2 (bases 1 to 3233) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3233 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 364..386 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 378..395 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 379..395 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1012..1028) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1012..1028) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1025..1047) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1036..1052 /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)." promoter complement(1060..1090) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1105..1126 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1243..1260) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1414..2002) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1494..1513) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2173..3033) /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 2796..2815 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3034..3138) /gene="bla" /label=AmpR promoter primer_bind 3206..3224 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 361 tttcccagtc acgacgttgt aaaacgacgg ccagtttcga actgcagttg gtacccgaat 421 tcgagctcct gaggtgtctc acaagtgccg tgcagtcccg cccccacttg cttctctttg 481 tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc cacctcgatc cggtctagac 541 tgaggtgtct cacaagtgcc gtgcagtccc gcccccactt gcttctcttt gtgtgtagtg 601 tacgtacatt atcgagaccg ttgttcccgc ccacctcgat ccggggatcc ctgaggtgtc 661 tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt tgtgtgtagt gtacgtacat 721 tatcgagacc gttgttcccg cccacctcga tccgggtcga cctgaggtgt ctcacaagtg 781 ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag tgtacgtaca ttatcgagac 841 cgttgttccc gcccacctcg atccgggcat gcactgatca cgggcaaaag tgcgtatata 901 tacaagagcg tttgccagcc acagattttc actccacaca ccacatcaca catacaacca 961 cacacatcca caatggaacc cgaaactaaa agcttcaggc gcgccatcat ggtcatagct 1021 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 1081 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 1141 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 1201 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 1261 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 1321 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 1381 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 1441 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 1501 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 1561 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 1621 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 1681 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 1741 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 1801 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 1861 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 1921 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 1981 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 2041 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 2101 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 2161 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 2221 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 2281 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 2341 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 2401 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 2461 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 2521 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 2581 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 2641 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 2701 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 2761 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 2821 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 2881 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 2941 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 3001 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 3061 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 3121 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 3181 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtc //