LOCUS Exported 5035 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pUC-UAS1B20-Leum SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5035) 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 5035) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5035 /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(2814..2830) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2814..2830) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2827..2849) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2838..2854 /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(2862..2892) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2907..2928 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3045..3062) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3216..3804) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3296..3315) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3975..4835) /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 4598..4617 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4836..4940) /gene="bla" /label=AmpR promoter primer_bind 5008..5026 /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 aggtacccga attcctgagg 421 tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc tctttgtgtg tagtgtacgt 481 acattatcga gaccgttgtt cccgcccacc tcgatccggt ctagactgag gtgtctcaca 541 agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg tacattatcg 601 agaccgttgt tcccgcccac ctcgatccgg ggatccctga ggtgtctcac aagtgccgtg 661 cagtcccgcc cccacttgct tctctttgtg tgtagtgtac gtacattatc gagaccgttg 721 ttcccgccca cctcgatccg ggtcgacctg aggtgtctca caagtgccgt gcagtcccgc 781 ccccacttgc ttctctttgt gtgtagtgta cgtacattat cgagaccgtt gttcccgccc 841 acctcgatcc gggagctcct gaggtgtctc acaagtgccg tgcagtcccg cccccacttg 901 cttctctttg tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc cacctcgatc 961 cggtctagac tgaggtgtct cacaagtgcc gtgcagtccc gcccccactt gcttctcttt 1021 gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc ccacctcgat ccggggatcc 1081 ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt tgtgtgtagt 1141 gtacgtacat tatcgagacc gttgttcccg cccacctcga tccgggtcga cctgaggtgt 1201 ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag tgtacgtaca 1261 ttatcgagac cgttgttccc gcccacctcg atccgggcat gcggtacccg aattcctgag 1321 gtgtctcaca agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt gtagtgtacg 1381 tacattatcg agaccgttgt tcccgcccac ctcgatccgg tctagactga ggtgtctcac 1441 aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac gtacattatc 1501 gagaccgttg ttcccgccca cctcgatccg gggatccctg aggtgtctca caagtgccgt 1561 gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat cgagaccgtt 1621 gttcccgccc acctcgatcc gggtcgacct gaggtgtctc acaagtgccg tgcagtcccg 1681 cccccacttg cttctctttg tgtgtagtgt acgtacatta tcgagaccgt tgttcccgcc 1741 cacctcgatc cgggagctcc tgaggtgtct cacaagtgcc gtgcagtccc gcccccactt 1801 gcttctcttt gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc ccacctcgat 1861 ccggtctaga ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt 1921 tgtgtgtagt gtacgtacat tatcgagacc gttgttcccg cccacctcga tccggggatc 1981 cctgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag 2041 tgtacgtaca ttatcgagac cgttgttccc gcccacctcg atccgggtcg acctgaggtg 2101 tctcacaagt gccgtgcagt cccgccccca cttgcttctc tttgtgtgta gtgtacgtac 2161 attatcgaga ccgttgttcc cgcccacctc gatccgggca tgcctgcagt tggtacccga 2221 attcgagctc ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt 2281 tgtgtgtagt gtacgtacat tatcgagacc gttgttcccg cccacctcga tccggtctag 2341 actgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag 2401 tgtacgtaca ttatcgagac cgttgttccc gcccacctcg atccggggat ccctgaggtg 2461 tctcacaagt gccgtgcagt cccgccccca cttgcttctc tttgtgtgta gtgtacgtac 2521 attatcgaga ccgttgttcc cgcccacctc gatccgggtc gacctgaggt gtctcacaag 2581 tgccgtgcag tcccgccccc acttgcttct ctttgtgtgt agtgtacgta cattatcgag 2641 accgttgttc ccgcccacct cgatccgggc atgcactgat cacgggcaaa agtgcgtata 2701 tatacaagag cgtttgccag ccacagattt tcactccaca caccacatca cacatacaac 2761 cacacacatc cacaatggaa cccgaaacta aaagcttcag gcgcgccatc atggtcatag 2821 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 2881 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 2941 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 3001 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 3061 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 3121 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 3181 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 3241 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 3301 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 3361 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 3421 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 3481 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 3541 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 3601 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 3661 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 3721 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 3781 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 3841 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 3901 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 3961 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 4021 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 4081 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 4141 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 4201 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 4261 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 4321 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 4381 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 4441 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 4501 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 4561 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 4621 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 4681 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 4741 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 4801 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 4861 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 4921 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc 4981 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc //