LOCUS       Exported                5891 bp ds-DNA     circular SYN 06-JUN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5891)
  AUTHORS   Hampton lab
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2019 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5891
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     184..206
                     /label=oRH5049
     promoter        187..420
                     /label=MET17 Promoter
     CDS             421..1755
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     primer_bind     complement(1753..1774)
                     /label=oRH5050
     rep_origin      complement(1860..2315)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1974..1993)
                     /label=oRH4736
     CDS             complement(2098..2676)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     2460..2476
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2486..2504
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2513..2620
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2530..2546
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2580..2596)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2633..2651)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2672..2688)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2696..2712
                     /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(2720..2750)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(3074..3662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3833..4693)
                     /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"
     promoter        complement(4694..4798)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4791..4850
                     /label=oRH4868
     promoter        4839..5119
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5120..5794
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(5823..5882)
                     /label=oRH4869
     primer_bind     5872..5891
                     /label=oRH4872
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatagcca tcctcatgaa aactgtgtaa cataataacc gaagtgtcga aaaggtggca
      241 ccttgtccaa ttgaacacgc tcgatgaaaa aaataagata tatataaggt taagtaaagc
      301 gtctgttaga aaggaagttt ttcctttttc ttgctctctt gtcttttcat ctactatttc
      361 cttcgtgtaa tacagggtcg tcagatacat agatacaatt ctattacccc catccataca
      421 atgccatctc atttcgatac tgttcaacta cacgccggcc aagagaaccc tggtgacaat
      481 gctcacagat ccagagctgt accaatttac gccaccactt cttatgtttt cgaaaactct
      541 aagcatggtt cgcaattgtt tggtctagaa gttccaggtt acgtctattc ccgtttccaa
      601 aacccaacca gtaatgtttt ggaagaaaga attgctgctt tagaaggtgg tgctgctgct
      661 ttggctgttt cctccggtca agccgctcaa acccttgcca tccaaggttt ggcacacact
      721 ggtgacaaca tcgtttccac ttcttactta tacggtggta cttataacca gttcaaaatc
      781 tcgttcaaaa gatttggtat cgaggctaga tttgttgaag gtgacaatcc agaagaattc
      841 gaaaaggtct ttgatgaaag aaccaaggct gtttatttgg aaaccattgg taatccaaag
      901 tacaatgttc cggattttga aaaaattgtt gcaattgctc acaaacacgg tattccagtt
      961 gtcgttgaca acacatttgg tgccggtggt tacttctgtc agccaattaa atacggtgct
     1021 gatattgtaa cacattctgc taccaaatgg attggtggtc atggtactac tatcggtggt
     1081 attattgttg actctggtaa gttcccatgg aaggactacc cagaaaagtt ccctcaattc
     1141 tctcaacctg ccgaaggata tcacggtact atctacaatg aagcctacgg taacttggca
     1201 tacatcgttc atgttagaac tgaactatta agagatttgg gtccattgat gaacccattt
     1261 gcctctttct tgctactaca aggtgttgaa acattatctt tgagagctga aagacacggt
     1321 gaaaatgcat tgaagttagc caaatggtta gaacaatccc catacgtatc ttgggtttca
     1381 taccctggtt tagcatctca ttctcatcat gaaaatgcta agaagtatct atctaacggt
     1441 ttcggtggtg tcttatcttt cggtgtaaaa gacttaccaa atgccgacaa ggaaactgac
     1501 ccattcaaac tttctggtgc tcaagttgtt gacaatttaa agcttgcctc taacttggcc
     1561 aatgttggtg atgccaagac cttagtcatt gctccatact tcactaccca caaacaatta
     1621 aatgacaaag aaaagttggc atctggtgtt accaaggact taattcgtgt ctctgttggt
     1681 atcgaattta ttgatgacat tattgcagac ttccagcaat cttttgaaac tgttttcgct
     1741 ggccaaaaac catgagtgtg cgtaatgagt tgtaaaatta tgtataaacc tactttctct
     1801 cacaagttat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     1861 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     1921 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     1981 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2041 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2101 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2161 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2221 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2281 acccgccgcg cttaatgcgc cgctacaggg cgcgtcgcgc cattcgccat tcaggctgcg
     2341 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     2401 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     2461 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
     2521 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg
     2581 atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
     2641 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2701 atccgctcac aattccacac aacataggag ccggaagcat aaagtgtaaa gcctggggtg
     2761 cctaatgagt gaggtaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2821 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2881 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2941 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3001 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3061 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3121 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3181 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3241 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3301 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3361 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3421 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3481 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     3541 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3601 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3661 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3721 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3781 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3841 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3901 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3961 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4021 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4081 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4141 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4201 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4261 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4321 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4381 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4441 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4501 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4561 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4621 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4681 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4741 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4801 catttccccg aaaagtgcca cctgacgtct gcaccataaa cgacattact atatatataa
     4861 tataggaagc atttaataga cagcatcgta atatatgtgt actttgcagt tatgacgcca
     4921 gatggcagta gtggaagata ttctttattg aaaaatagct tgtcacctta cgtacaatct
     4981 tgatccggag cttttctttt tttgccgatt aagaattaat tcggtcgaaa aaagaaaagg
     5041 agagggccaa gagggagggc attggtgact attgagcacg tgagtatacg tgattaagca
     5101 cacaaaggca gcttggagta tgtctgttat taatttcaca ggtagttctg gtccattggt
     5161 gaaagtttgc ggcttgcaga gcacagaggc cgcagaatgt gctctagatt ccgatgctga
     5221 cttgctgggt attatatgtg tgcccaatag aaagagaaca attgacccgg ttattgcaag
     5281 gaaaatttca agtcttgtaa aagcatataa aaatagttca ggcactccga aatacttggt
     5341 tggcgtgttt cgtaatcaac ctaaggagga tgttttggct ctggtcaatg attacggcat
     5401 tgatatcgtc caactgcatg gagatgagtc gtggcaagaa taccaagagt tcctcggttt
     5461 gccagttatt aaaagactcg tatttccaaa agactgcaac atactactca gtgcagcttc
     5521 acagaaacct cattcgttta ttcccttgtt tgattcagaa gcaggtggga caggtgaact
     5581 tttggattgg aactcgattt ctgactgggt tggaaggcaa gagagccccg aaagcttaca
     5641 ttttatgtta gctggtggac tgacgccaga aaatgttggt gatgcgctta gattaaatgg
     5701 cgttattggt gttgatgtaa gcggaggtgt ggagacaaat ggtgtaaaag actctaacaa
     5761 aatagcaaat ttcgtcaaaa atgctaagaa ataggttatt actgagtagt atttatttaa
     5821 gtattgtttg tgcacttgcc taatcatgac attaacctat aaaaataggc gtatcacgag
     5881 gccctttcgt c
//
