LOCUS       40924_49532        7769 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast episomal cloning vector with a URA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7769)
  AUTHORS   New England Biolabs
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7769)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     See also GenBank accession L09156.
FEATURES             Location/Qualifiers
     source          1..7769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      344..1686
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        2278..2498
                     /label=URA3 promoter
     CDS             2499..3299
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             3495..4682
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             5326..5514
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    5619..5759
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5945..6533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6707..7564)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7565..7669)
                     /label=AmpR promoter
ORIGIN
        1 gaattctgaa ccagtcctaa aacgagtaaa taggaccggc aattcttcaa gcaataaaca
       61 ggaataccaa ttattaaaag ataacttagt cagatcgtac aataaagctt tgaagaaaaa
      121 tgcgccttat tcaatctttg ctataaaaaa tggcccaaaa tctcacattg gaagacattt
      181 gatgacctca tttctttcaa tgaagggcct aacggagttg actaatgttg tgggaaattg
      241 gagcgataag cgtgcttctg ccgtggccag gacaacgtat actcatcaga taacagcaat
      301 acctgatcac tacttcgcac tagtttctcg gtactatgca tatgatccaa tatcaaagga
      361 aatgatagca ttgaaggatg agactaatcc aattgaggag tggcagcata tagaacagct
      421 aaagggtagt gctgaaggaa gcatacgata ccccgcatgg aatgggataa tatcacagga
      481 ggtactagac tacctttcat cctacataaa tagacgcata taagtacgca tttaagcata
      541 aacacgcact atgccgttct tctcatgtat atatatatac aggcaacacg cagatatagg
      601 tgcgacgtga acagtgagct gtatgtgcgc agctcgcgtt gcattttcgg aagcgctcgt
      661 tttcggaaac gctttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac
      721 ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa aaccaaaaac
      781 gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa acattgctca
      841 aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac ccatccacct
      901 ttcgctcctt gaacttgcat ctaaactcga cctctacatt ttttatgttt atctctagta
      961 ttactcttta gacaaaaaaa ttgtagtaag aactattcat agagtgaatc gaaaacaata
     1021 cgaaaatgta aacatttcct atacgtagta tatagagaca aaatagaaga aaccgttcat
     1081 aattttctga ccaatgaaga atcatcaacg ctatcacttt ctgttcacaa agtatgcgca
     1141 atccacatcg gtatagaata taatcgggga tgcctttatc ttgaaaaaat gcacccgcag
     1201 cttcgctagt aatcagtaaa cgcgggaagt ggagtcaggc tttttttatg gaagagaaaa
     1261 tagacaccaa agtagccttc ttctaacctt aacggaccta cagtgcaaaa agttatcaag
     1321 agactgcatt atagagcgca caaaggagaa aaaaagtaat ctaagatgct ttgttagaaa
     1381 aatagcgctc tcgggatgca tttttgtaga acaaaaaaga agtatagatt ctttgttggt
     1441 aaaatagcgc tctcgcgttg catttctgtt ctgtaaaaat gcagctcaga ttctttgttt
     1501 gaaaaattag cgctctcgcg ttgcattttt gttttacaaa aatgaagcac agattcttcg
     1561 ttggtaaaat agcgctttcg cgttgcattt ctgttctgta aaaatgcagc tcagattctt
     1621 tgtttgaaaa attagcgctc tcgcgttgca tttttgttct acaaaatgaa gcacagatgc
     1681 ttcgttaaca aagatatgct attgaagtgc aagatggaaa cgcagaaaat gaaccgggga
     1741 tgcgacgtgc aagattacct atgcaataga tgcaatagtt tctccaggaa ccgaaataca
     1801 tacattgtct tccgtaaagc gctagactat atattattat acaggttcaa atatactatc
     1861 tgtttcaggg aaaactccca ggttcggatg ttcaaaattc aatgatgggt aacaagtacg
     1921 atcgtaaatc tgtaaaacag tttgtcggat attaggctgt atctcctcaa agcgtattcg
     1981 aatatcattg agaagctgca gcgtcacatc ggataataat gatggcagcc attgtagaag
     2041 tgccttttgc atttctagtc tctttctcgg tctagctagt tttactacat cgcgaagata
     2101 gaatcttaga tcacactgcc tttgctgagc tggatcaata gagtaacaaa agagtggtaa
     2161 ggcctcgtta aaggacaagg acctgagcgg aagtgtatcg tacagtagac ggagtatact
     2221 agtatagtct atagtccgtg gaattctcat gtttgacagc ttatcatcga taagcttttc
     2281 aattcaattc atcatttttt ttttattctt ttttttgatt tcggtttctt tgaaattttt
     2341 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     2401 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     2461 tgcacagaac aaaaacctgc aggaaacgaa gataaatcat gtcgaaagct acatataagg
     2521 aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc atgcacgaaa
     2581 agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta ctggagttag
     2641 ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc ttgactgatt
     2701 tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac aattttttac
     2761 tcttcgaaga cagaaaattt gctgacattg gtaatacagt caaattgcag tactctgcgg
     2821 gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg gtgggcccag
     2881 gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct agaggccttt
     2941 tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat actaagggta
     3001 ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct caaagagaca
     3061 tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg ggtttagatg
     3121 acaagggaga cgcattgggt caacagtata gaaccgtgga tgatgtggtc tctacaggat
     3181 ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct aaggtagagg
     3241 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact
     3301 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
     3361 aattatatca gttattaccc gggaatctcg gtcgtaatga tttttataat gacgaaaaaa
     3421 aaaaaattgg aaagaaaaag ctttaatgcg gtagtttatc acagttaaat tgctaacgca
     3481 gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt catcctcggc accgtcaccc
     3541 tggatgctgt aggcataggc ttggttatgc cggtactgcc gggcctcttg cgggatatcg
     3601 tccattccga cagcatcgcc agtcactatg gcgtgctgct agcgctatat gcgttgatgc
     3661 aatttctatg cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc
     3721 tgctcgcttc gctacttgga gccactatcg actacgcgat catggcgacc acacccgtcc
     3781 tgtggatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
     3841 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     3901 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
     3961 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac
     4021 tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgaccgatg cccttgagag
     4081 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     4141 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
     4201 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
     4261 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
     4321 agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt
     4381 tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg
     4441 ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc
     4501 ttcaaggatc gctcgcggct cttaccagcc taacttcgat cactggaccg ctgatcgtca
     4561 cggcgattta tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg
     4621 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     4681 cctgaatgga agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa
     4741 tcaattcttg cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc
     4801 gtccgccatc tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg
     4861 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     4921 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     4981 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     5041 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
     5101 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
     5161 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
     5221 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
     5281 cattaccccc atgaacagaa attccccctt acacggaggc atcaagtgac caaacaggaa
     5341 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     5401 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     5461 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5521 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5581 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     5641 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     5701 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     5761 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5821 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5881 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6121 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6301 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6361 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6421 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6481 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg
     7021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7321 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7681 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7741 ggcgtatcac gaggcccttt cgtcttcaa
//
