LOCUS Exported 4520 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRSII408 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4520) 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 4520) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4520 /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(1511..1966) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1648..1669) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1860..1879 /label=F1ori-R /note="F1 origin, reverse primer" CDS complement(1943..2323) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR TDRPSQQLRSLNGEWTRPVAAH" primer_bind 2092..2114 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2106..2123 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2107..2123 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2133..2152 /label=T7 /note="T7 promoter, forward primer" promoter 2133..2151 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 2160..2267 /label=MCS /note="pBluescript multiple cloning site" primer_bind 2176..2192 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 2177..2193 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2227..2243) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2227..2243) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2280..2300) /label=T3 /note="T3 promoter, forward primer" promoter complement(2280..2298) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2319..2335) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2319..2335) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2332..2354) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2343..2359 /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(2367..2397) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2412..2433 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2550..2567) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2721..3309) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2801..2820) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3480..4340) /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 4103..4122 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4341..4445) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatagatc tcttattcac gagtcagtct gactcttgcg agagatgagg atgtaataat 241 actaatctcg aagatgccat ctaatacata tagacataca tatatatata tatacattct 301 atatattctt acccagattc tttgaggtaa gacggttggg ttttatcttt tgcagttggt 361 actattaaga acaatcgaat cataagcatt gcttacaaag aatacacata cgaaatatta 421 acgataatgt caattacgaa gactgaactg gacggtatat tgccattggt ggccagaggt 481 aaagttagag acatatatga ggtagacgct ggtacgttgc tgtttgttgc tacggatcgt 541 atctctgcat atgacgttat tatggaaaac agcattcctg aaaagggtat cctattgacc 601 aaactgtcag agttctggtt caagttcctg tccaacgatg ttcgtaatca tttggtagac 661 atcgccccag gtaagactat tttcgattat ctacctgcaa aattgagcga accaaagtac 721 aaaacgcaac tagaagaccg ctctctattg gttcacaaac ataaactaat tccattggaa 781 gtaattgtca gaggctacat caccggatct gcttggaaag agtacgtaaa aacaggtact 841 gtgcatggtt tgaaacaacc tcaaggactt aaagaatctc aagagttccc agaaccaatc 901 ttcaccccat cgaccaaggc tgaacaaggt gaacatgacg aaaacatctc tcctgcccag 961 gccgctgagc tggtgggtga agatttgtca cgtagagtgg cagaactggc tgtaaaactg 1021 tactccaagt gcaaagatta tgctaaggag aagggcatca tcatcgcaga cactaaattc 1081 gaatttggta ttgacgaaaa gaccaatgaa attattctag tggacgaggt gctaacgcca 1141 gactcctcaa gattctggaa cggtgcctct tataaggtag gagaatccca agattcttac 1201 gataagcaat ttttaagaga ctggcttact gctaataagt tgaacggtgt taacggcgtc 1261 aaaatgcccc aagacattgt agacaggaca agggccaaat atatagaggc ttatgaaaca 1321 ttgacagggt ctaaatggtc tcactaacgt gatttacata tactacaagt cgccagtgta 1381 actcctcact gaatatgatt catacatacc cgtatgtatt aatgtataaa tgttctcaga 1441 gcaaatttta tcgagatcta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 1501 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa 1561 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa 1621 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac 1681 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa 1741 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct 1801 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa 1861 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc 1921 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca 1981 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg 2041 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac 2101 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg 2161 gtaccgggcc ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc 2221 ccgggggatc cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc 2281 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga 2341 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 2401 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 2461 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 2521 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 2581 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 2641 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 2701 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 2761 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 2821 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 2881 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 2941 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 3001 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 3061 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 3121 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 3181 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 3241 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 3301 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 3361 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 3421 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 3481 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 3541 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 3601 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 3661 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 3721 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 3781 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 3841 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 3901 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 3961 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 4021 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 4081 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 4141 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 4201 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 4261 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 4321 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 4381 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 4441 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 4501 ttaacctata aaaataggcg //