LOCUS Exported 4969 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRSII413 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4969) 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 4969) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4969 /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" promoter 317..504 /gene="S. cerevisiae HIS3" /label=HIS3 promoter CDS 505..1167 /codon_start=1 /gene="S. cerevisiae HIS3" /product="imidazoleglycerol-phosphate dehydratase, required for histidine biosynthesis" /label=HIS3 /note="yeast auxotrophic marker" /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM" rep_origin complement(1427..1882) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1564..1585) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1776..1795 /label=F1ori-R /note="F1 origin, reverse primer" CDS complement(1859..2239) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR TDRPSQQLRSLNGEWTRPVAAH" primer_bind 2008..2030 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2022..2039 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2023..2039 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2049..2068 /label=T7 /note="T7 promoter, forward primer" promoter 2049..2067 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 2076..2183 /label=MCS /note="pBluescript multiple cloning site" primer_bind 2092..2108 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 2093..2109 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2143..2159) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2143..2159) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2196..2216) /label=T3 /note="T3 promoter, forward primer" promoter complement(2196..2214) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2235..2251) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2235..2251) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2248..2270) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2259..2275 /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(2283..2313) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2328..2349 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2466..2483) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2637..3225) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2717..2736) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3396..4256) /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 4019..4038 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4257..4361) /gene="bla" /label=AmpR promoter misc_feature 4398..4901 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" primer_bind 4942..4960 /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 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt 241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta 301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat 361 tttttttttt ccacctagcg gatgactctt tttttttctt agcgattggc attatcacat 421 aatgaattat acattatata aagtaatgtg atttcttcga agaatatact aaaaaatgag 481 caggcaagat aaacgaaggc aaagatgaca gagcagaaag ccctagtaaa gcgtattaca 541 aatgaaacca agattcagat tgcgatctct ttaaagggtg gtcccctagc gatagagcac 601 tcgatcttcc cagaaaaaga ggcagaagca gtagcagaac aggccacaca atcgcaagtg 661 attaacgtcc acacaggtat agggtttctg gaccatatga tacatgctct ggccaagcat 721 tccggctggt cgctaatcgt tgagtgcatt ggtgacttac acatagacga ccatcacacc 781 actgaagact gcgggattgc tctcggtcaa gcatttaaag aggccctagg ggccgtgcgt 841 ggagtaaaaa ggtttggatc aggatttgcg cctttggatg aggcactttc cagagcggtg 901 gtagatcttt cgaacaggcc gtacgcagtt gtcgaacttg gtttgcaaag ggagaaagta 961 ggagatctct cttgcgagat gatcccgcat tttcttgaaa gttttgcaga ggctagcaga 1021 attaccctcc acgttgattg tctgcgaggc aagaatgatc atcaccgtag tgagagtgcg 1081 ttcaaggctc ttgcggttgc cataagagaa gccacctcgc ccaatggtac aaacgatgtt 1141 ccctccacca aaggtgttct tatgtagtga caccgattat ttaaagttgc agcatacgat 1201 atatatacat gtgtatatat gtatacctat gaatgtcagt aagtatgtat acgaacagta 1261 tgatactgaa gatgacaagg taatgcatca ttctatacgt gtcattctga acgaggcgcg 1321 ctttcctttt ttctttttgc tttttctttt tttttctctt gaactcgacg gatctatgcg 1381 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt 1441 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag 1501 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt 1561 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga 1621 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg 1681 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct 1741 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc 1801 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt 1861 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc 1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 2041 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg 2101 acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact agttctagag 2161 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc 2221 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 2281 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 2341 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 2401 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 2461 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 2521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 2581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 2641 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 2701 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 2761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 2821 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 2881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 2941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 3001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 3061 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 3121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 3181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 3241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 3301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 3361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 3421 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 3481 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 3541 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 3601 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 3661 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 3721 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 3781 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 3841 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 3901 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 3961 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 4021 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 4081 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 4141 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 4201 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 4261 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 4321 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 4381 ccacctgggt ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat 4441 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt 4501 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc 4561 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca 4621 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct 4681 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc 4741 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa 4801 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca 4861 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtctaaga aaccattatt 4921 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc //