LOCUS Exported 5031 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRSII409 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5031) AUTHORS Chee MK, Haase SB TITLE New and Redesigned pRS Plasmid Shuttle Vectors for Genetic Manipulation of Saccharomycescerevisiae. JOURNAL G3 (Bethesda). 2012 May;2(5):515-26. Epub 2012 May 1. PUBMED 22670222 REFERENCE 2 (bases 1 to 5031) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5031 /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" rep_origin complement(2002..2457) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2139..2160) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2351..2370 /label=F1ori-R /note="F1 origin, reverse primer" CDS complement(2434..2814) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR TDRPSQQLRSLNGEWTRPVAAH" primer_bind 2583..2605 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2597..2614 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2598..2614 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2624..2643 /label=T7 /note="T7 promoter, forward primer" promoter 2624..2642 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 2651..2758 /label=MCS /note="pBluescript multiple cloning site" primer_bind 2667..2683 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 2668..2684 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2718..2734) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2718..2734) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2771..2791) /label=T3 /note="T3 promoter, forward primer" promoter complement(2771..2789) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2810..2826) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2810..2826) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2823..2845) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2834..2850 /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(2858..2888) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2903..2924 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3041..3058) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3212..3800) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3292..3311) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3971..4831) /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 4594..4613 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4832..4936) /gene="bla" /label=AmpR promoter primer_bind 5004..5022 /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 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc 241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag 301 acttagattg gtatatatac gcatatgcga gtcggcgagc aaacagggtg ctgctactaa 361 acgaacaata aagaaagaca atttgggcac ccaatataaa atgagcagtc aattcaatca 421 attcagtgtg aaattttttt cttggaaaga tcattcactt ctacacagct gtacatgagt 481 ttatatgaca gttttttttt atgtacatcc agttcaagat aatcagatta tatattggcc 541 caaaagggat cattaacgaa ttgcgaaatg ggtaatctct ttacagttaa cagattctct 601 ggtgattggc tttcctcaag gtagcaaata tactctaatt gtagcacgtc tactatgtat 661 ttctttacct tgtcatagca cacacccact tgcgccacgc cgtgtgcgtc atcacttaga 721 acaaatctgg agccacagtg cttcttgacc agattacata aggttttgct ggggtacggc 781 tcctccaggc gctttcttaa tgcggacgta ttgatttcaa ttgcgccgcc ataggagtct 841 ataaattgta aatttcttac aactgcatcg tatatttctg gccattcact gatgacgtcc 901 agtgaagcta caggaactcc ggtttcttcg ttgcagttgc ccgatttctg gtttactagc 961 atgtcattgg gcaaaaataa tttgtaaagg tcgaagtgac ccacgaccaa cggtttaata 1021 ttgatcagca tttcgtactg tgattggaag taagacagga gaaaatgttt caaattatca 1081 ttgaaggaat gcaatgaatt gtaccattgt tgttggtcga aatcaatagg tatcccgttg 1141 acgtgatgga ccgaacccac acaaaacttc aaaatatcat tattctcctt catgagtcgc 1201 tttgcatatt cgatatgagc catgtcacaa ctttcgatct ccattcctat aatgaattta 1261 gtccgcacat cgggtctatc agcataacga gtcttgattt cttgcgcatg actcatgaaa 1321 ttcttgaacg atgtttctag cttggttatg acttcctcag gattcttgcc caatgactgc 1381 tcttcggggt atataaactt ggcctcaatt cttggtatgt gctctgtcaa acagtacgtg 1441 tgaaagttga ggttgaccac ttgatcgacc acggaatcca aagggtccgt accgtgggca 1501 ctatagtcac cggagtgtga atggtgtgag tgcatgacct tgaggatttt tactgaattg 1561 tacgtatcaa gaagtactaa aatgtgtttg aaagggaaag attgtacggg aaatggaaaa 1621 tgtacatata atatccagta tcatttgagt tgatagagat tatctctata gatagagaga 1681 tttgacaaaa aaaataaaaa aatttaaatg gacgagtccg gaatcgaacc ggagacctct 1741 cccatgctaa gggagcgcgc taccgactac gccacacgcc cgaacgtttg ttgaaatctt 1801 ccaaacagaa atgatacagg tttactttac agcaagacat tagtagtttc attagcttat 1861 caacatgtat gaaaccggca agattgaaag atgcatgtat actaaactca caaattagag 1921 cttcaattta attatatcag ttattaccct atgcggtgtg aaataccgca cagatgcgta 1981 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa 2041 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata 2101 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac 2161 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc 2221 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa 2281 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg 2341 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg 2401 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca 2461 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt 2521 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt 2581 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata 2641 gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga 2701 attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca 2761 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt 2821 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 2881 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 2941 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 3001 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 3061 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 3121 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 3181 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 3241 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 3301 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 3361 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 3421 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 3481 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 3541 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 3601 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 3661 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 3721 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 3781 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 3841 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 3901 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 3961 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 4021 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 4081 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 4141 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 4201 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 4261 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 4321 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 4381 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 4441 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 4501 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 4561 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 4621 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 4681 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 4741 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 4801 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 4861 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 4921 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta 4981 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt c //