LOCUS Exported 3560 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pUC-UAS1B7-Leum SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3560) 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 3560) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3560 /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(1339..1355) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1339..1355) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1352..1374) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1363..1379 /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(1387..1417) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1432..1453 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1570..1587) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1741..2329) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1821..1840) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2500..3360) /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 3123..3142 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3361..3465) /gene="bla" /label=AmpR promoter primer_bind 3533..3551 /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 tcctgaggtg tctcacaagt gccgtgcagt cccgccccca cttgcttctc tttgtgtgta 481 gtgtacgtac attatcgaga ccgttgttcc cgcccacctc gatccggtct agactgaggt 541 gtctcacaag tgccgtgcag tcccgccccc acttgcttct ctttgtgtgt agtgtacgta 601 cattatcgag accgttgttc ccgcccacct cgatccgggg atccctgagg tgtctcacaa 661 gtgccgtgca gtcccgcccc cacttgcttc tctttgtgtg tagtgtacgt acattatcga 721 gaccgttgtt cccgcccacc tcgatccggg tcgacctgag gtgtctcaca agtgccgtgc 781 agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg tacattatcg agaccgttgt 841 tcccgcccac ctcgatccgg gagctcctga ggtgtctcac aagtgccgtg cagtcccgcc 901 cccacttgct tctctttgtg tgtagtgtac gtacattatc gagaccgttg ttcccgccca 961 cctcgatccg gggatccctg aggtgtctca caagtgccgt gcagtcccgc ccccacttgc 1021 ttctctttgt gtgtagtgta cgtacattat cgagaccgtt gttcccgccc acctcgatcc 1081 gggtcgacct gaggtgtctc acaagtgccg tgcagtcccg cccccacttg cttctctttg 1141 tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc cacctcgatc cgggcatgca 1201 ctgatcacgg gcaaaagtgc gtatatatac aagagcgttt gccagccaca gattttcact 1261 ccacacacca catcacacat acaaccacac acatccacaa tggaacccga aactaaaagc 1321 ttcaggcgcg ccatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 1381 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 1441 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 1501 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 1561 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 1621 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 1681 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 1741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 1801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 1861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 1921 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 1981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 2041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 2101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 2161 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 2221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 2281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 2341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 2401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 2461 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 2521 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 2581 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 2641 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 2701 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 2761 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 2821 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 2881 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 2941 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 3001 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 3061 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 3121 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 3181 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 3241 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 3301 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 3361 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 3421 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 3481 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 3541 tatcacgagg ccctttcgtc //