LOCUS pRS316 4887 bp ds-DNA circular SYN 09-九月-2012
DEFINITION Yeast centromere vector with URA3 marker.
ACCESSION U03442
VERSION .
KEYWORDS pRS316
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4887)
AUTHORS Sikorski RS, Hieter P.
TITLE A system of shuttle vectors and yeast host strains designed for
efficient manipulation of DNA in Saccharomyces cerevisiae.
JOURNAL Genetics 1989;122:19-27.
PUBMED 2659436
REFERENCE 2 (bases 1 to 4887)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2013年8月13日 from SnapGene Viewer 1.5.3
http://www.snapgene.com
COMMENT
FEATURES Location/Qualifiers
source 1..4887
/organism="synthetic DNA construct"
/lab_host="Saccharomyces cerevisiae"
/mol_type="other DNA"
promoter 190..410
/gene="S. cerevisiae URA3"
/note="URA3 promoter"
/note="color: #ffffff; direction: RIGHT"
CDS 411..1214
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/note="URA3"
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/note="color: #ff7f50"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
rep_origin complement(1343..1798)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
/note="color: #ffff00"
CDS complement(1581..2156)
/codon_start=1
/gene="lacZ?fragment"
/product="fragment of?beta-galactosidase"
/note="lacZ'"
/note="color: #993366"
/translation="MTMITPSSELTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
LERPPPRWSSNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
FPRQALNRGLPLGFRFSALRHLDPKKLD"
primer_bind 1943..1959
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
/note="color: #a020f0; direction: RIGHT"
promoter 1966..1984
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
/note="color: #ffffff; direction: RIGHT"
misc_feature 1993..2100
/note="MCS"
/note="pBluescript multiple cloning site"
/note="color: #99ccff"
promoter complement(2113..2131)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
/note="color: #ffffff; direction: LEFT"
primer_bind complement(2152..2168)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
/note="color: #a020f0; direction: LEFT"
protein_bind 2176..2192
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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)."
/note="color: #31849b"
promoter complement(2200..2230)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
/note="This reverse directional feature has 3 segments:
???1:?2200?..?2206?/?#ffffff?/?-10
???2:?2207?..?2224?/?#ffffff
???3:?2225?..?2230?/?#ffffff?/?-35"
rep_origin complement(2554..3142)
/direction=LEFT
/note="ori"
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(3313..4173)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/note="This feature has 2 segments:
???1:?3313?..?4104?/?#ccffcc
???2:?4105?..?4173?/?#ccffcc?/?signal?sequence
Cleavage site after base 4104"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4174..4278)
/gene="bla"
/note="AmpR promoter"
/note="color: #ffffff; direction: LEFT"
misc_feature 4315..4818
/note="CEN/ARS"
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
/note="color: #454f59"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
1981 tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtgg atcccccggg
2041 ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg gcccggtacc
2101 cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct
2161 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat
2221 aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc
2281 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
2341 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
2401 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
2461 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
2521 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctcggcc cccctgacga
2581 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
2641 ccaggcgttc ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
2701 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
2761 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
2821 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
2881 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
2941 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
3001 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
3061 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
3121 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
3181 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
3241 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
3301 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
3361 tcgttcatcc atagttgcct gactgcccgt cgtgtagata actacgatac gggagggctt
3421 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
3481 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
3541 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
3601 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
3661 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
3721 gtgaaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
3781 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
3841 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
3901 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
3961 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
4021 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
4081 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
4141 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
4201 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
4261 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg
4321 tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa
4381 agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta
4441 gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc
4501 cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt
4561 ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata
4621 tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat
4681 tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat
4741 aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta
4801 agttacaggc aagcgatccg tcctaagaaa ccattattat catgacatta acctataaaa
4861 ataggcgtat cacgaggccc tttcgtc
//