LOCUS Exported 3901 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pUC-UAS1B10-Leum SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3901) 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 3901) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3901 /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(1680..1696) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1680..1696) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1693..1715) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1704..1720 /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(1728..1758) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1773..1794 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1911..1928) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2082..2670) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2162..2181) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2841..3701) /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 3464..3483 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3702..3806) /gene="bla" /label=AmpR promoter primer_bind 3874..3892 /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 ggggatccct 1321 gaggtgtctc acaagtgccg tgcagtcccg cccccacttg cttctctttg tgtgtagtgt 1381 acgtacatta tcgagaccgt tgttcccgcc cacctcgatc cgggtcgacc tgaggtgtct 1441 cacaagtgcc gtgcagtccc gcccccactt gcttctcttt gtgtgtagtg tacgtacatt 1501 atcgagaccg ttgttcccgc ccacctcgat ccgggcatgc actgatcacg ggcaaaagtg 1561 cgtatatata caagagcgtt tgccagccac agattttcac tccacacacc acatcacaca 1621 tacaaccaca cacatccaca atggaacccg aaactaaaag cttcaggcgc gccatcatgg 1681 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 1741 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 1801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 1861 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 1921 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 1981 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 2041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 2101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 2161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 2221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 2281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 2341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 2401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 2461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 2521 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 2581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 2641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 2701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 2761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 2821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 2881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 2941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 3001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 3061 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 3121 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 3181 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 3241 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 3301 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 3361 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 3421 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 3481 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 3541 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 3601 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 3661 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 3721 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 3781 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa 3841 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt 3901 c //