LOCUS Exported 4454 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pUC-UAS1B15-Leum SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4454) 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 4454) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4454 /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(2233..2249) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2233..2249) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2246..2268) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2257..2273 /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(2281..2311) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2326..2347 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2464..2481) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2635..3223) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2715..2734) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3394..4254) /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 4017..4036 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4255..4359) /gene="bla" /label=AmpR promoter primer_bind 4427..4445 /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 tccggggatc cctgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct 1921 ttgtgtgtag tgtacgtaca ttatcgagac cgttgttccc gcccacctcg atccgggtcg 1981 acctgaggtg tctcacaagt gccgtgcagt cccgccccca cttgcttctc tttgtgtgta 2041 gtgtacgtac attatcgaga ccgttgttcc cgcccacctc gatccgggca tgcactgatc 2101 acgggcaaaa gtgcgtatat atacaagagc gtttgccagc cacagatttt cactccacac 2161 accacatcac acatacaacc acacacatcc acaatggaac ccgaaactaa aagcttcagg 2221 cgcgccatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 2281 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 2341 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 2401 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 2461 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 2521 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 2581 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 2641 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 2701 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 2761 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 2821 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 2881 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 2941 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 3001 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 3061 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 3121 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 3181 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 3241 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 3301 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 3361 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 3421 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 3481 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 3541 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 3601 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 3661 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 3721 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 3781 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 3841 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 3901 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 3961 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 4021 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 4081 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 4141 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 4201 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 4261 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 4321 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 4381 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 4441 gaggcccttt cgtc //