LOCUS Exported 3132 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pUC-UAS1B3-Leum SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3132) 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 3132) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3132 /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(911..927) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(911..927) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(924..946) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 935..951 /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(959..989) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1004..1025 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1142..1159) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1313..1901) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1393..1412) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2072..2932) /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 2695..2714 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2933..3037) /gene="bla" /label=AmpR promoter primer_bind 3105..3123 /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 agtctagact gaggtgtctc acaagtgccg tgcagtcccg cccccacttg 481 cttctctttg tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc cacctcgatc 541 cggggatccc tgaggtgtct cacaagtgcc gtgcagtccc gcccccactt gcttctcttt 601 gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc ccacctcgat ccgggtcgac 661 ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt tgtgtgtagt 721 gtacgtacat tatcgagacc gttgttcccg cccacctcga tccgggcatg cactgatcac 781 gggcaaaagt gcgtatatat acaagagcgt ttgccagcca cagattttca ctccacacac 841 cacatcacac atacaaccac acacatccac aatggaaccc gaaactaaaa gcttcaggcg 901 cgccatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 961 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 1021 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 1081 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 1141 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 1201 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 1261 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 1321 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 1381 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 1441 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 1501 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 1561 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 1621 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 1681 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 1741 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 1801 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 1861 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 1921 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 1981 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 2041 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 2101 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 2161 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 2221 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 2281 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 2341 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 2401 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 2461 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 2521 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 2581 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 2641 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 2701 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 2761 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 2821 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 2881 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 2941 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 3001 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 3061 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga 3121 ggccctttcg tc //