LOCUS Exported 4232 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pUC-UAS1B13-Leum SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4232) 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 4232) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4232 /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(2011..2027) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2011..2027) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2024..2046) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2035..2051 /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(2059..2089) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2104..2125 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2242..2259) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2413..3001) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2493..2512) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3172..4032) /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 3795..3814 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4033..4137) /gene="bla" /label=AmpR promoter primer_bind 4205..4223 /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 tccgggcatg cactgatcac gggcaaaagt gcgtatatat acaagagcgt ttgccagcca 1921 cagattttca ctccacacac cacatcacac atacaaccac acacatccac aatggaaccc 1981 gaaactaaaa gcttcaggcg cgccatcatg gtcatagctg tttcctgtgt gaaattgtta 2041 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 2101 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 2161 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 2221 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 2281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 2341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 2401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 2461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 2521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 2581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 2641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 2701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 2761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 2821 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 2881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 2941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 3001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 3061 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 3121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 3181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 3241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 3301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 3361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 3421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 3481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 3541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 3601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 3661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 3721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 3781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 3841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 3901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 3961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 4021 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 4081 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 4141 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 4201 taaaaatagg cgtatcacga ggccctttcg tc //