LOCUS Exported 4343 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pUC-UAS1B14-Leum SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4343) 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 4343) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4343 /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(2122..2138) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2122..2138) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2135..2157) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2146..2162 /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(2170..2200) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2215..2236 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2353..2370) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2524..3112) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2604..2623) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3283..4143) /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 3906..3925 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4144..4248) /gene="bla" /label=AmpR promoter primer_bind 4316..4334 /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 aattcctgag 421 gtgtctcaca agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg 481 tacattatcg agaccgttgt tcccgcccac ctcgatccgg tctagactga ggtgtctcac 541 aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac gtacattatc 601 gagaccgttg ttcccgccca cctcgatccg gggatccctg aggtgtctca caagtgccgt 661 gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat cgagaccgtt 721 gttcccgccc acctcgatcc gggtcgacct gaggtgtctc acaagtgccg tgcagtcccg 781 cccccacttg cttctctttg tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc 841 cacctcgatc cgggagctcc tgaggtgtct cacaagtgcc gtgcagtccc gcccccactt 901 gcttctcttt gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc ccacctcgat 961 ccggtctaga ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt 1021 tgtgtgtagt gtacgtacat tatcgagacc gttgttcccg cccacctcga tccggggatc 1081 cctgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag 1141 tgtacgtaca ttatcgagac cgttgttccc gcccacctcg atccgggtcg acctgaggtg 1201 tctcacaagt gccgtgcagt cccgccccca cttgcttctc tttgtgtgta gtgtacgtac 1261 attatcgaga ccgttgttcc cgcccacctc gatccgggca tgcggtaccc gaattcctga 1321 ggtgtctcac aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac 1381 gtacattatc gagaccgttg ttcccgccca cctcgatccg gtctagactg aggtgtctca 1441 caagtgccgt gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat 1501 cgagaccgtt gttcccgccc acctcgatcc ggggatccct gaggtgtctc acaagtgccg 1561 tgcagtcccg cccccacttg cttctctttg tgtgtagtgt acgtacatta tcgagaccgt 1621 tgttcccgcc cacctcgatc cgggtcgacc tgaggtgtct cacaagtgcc gtgcagtccc 1681 gcccccactt gcttctcttt gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc 1741 ccacctcgat ccgggagctc ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact 1801 tgcttctctt tgtgtgtagt gtacgtacat tatcgagacc gttgttcccg cccacctcga 1861 tccgggtcga cctgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct 1921 ttgtgtgtag tgtacgtaca ttatcgagac cgttgttccc gcccacctcg atccgggcat 1981 gcactgatca cgggcaaaag tgcgtatata tacaagagcg tttgccagcc acagattttc 2041 actccacaca ccacatcaca catacaacca cacacatcca caatggaacc cgaaactaaa 2101 agcttcaggc gcgccatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 2161 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 2221 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 2281 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 2341 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 2401 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 2461 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 2521 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 2581 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 2641 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 2701 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 2761 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 2821 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 2881 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 2941 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 3001 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 3061 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 3121 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 3181 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 3241 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 3301 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 3361 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 3421 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 3481 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 3541 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 3601 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 3661 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 3721 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 3781 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 3841 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 3901 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 3961 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 4021 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 4081 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 4141 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 4201 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 4261 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 4321 gcgtatcacg aggccctttc gtc //