LOCUS       Exported                7102 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 7102)
  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..7102
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        complement(35..53)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(74..90)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    98..114
                     /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(122..152)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(476..1064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1235..2095)
                     /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(2096..2200)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(2251..3585)
                     /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"
     promoter        complement(3586..3819)
                     /label=MET17 Promoter
     primer_bind     complement(3800..3847)
                     /label=oRH5049
     primer_bind     3850..3869
                     /label=oRH4872
     promoter        4068..4443
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             4444..6159
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     rep_origin      complement(6364..6819)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6478..6497)
                     /label=oRH4736
     CDS             complement(join(6602..7102,1..78))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     6964..6980
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6990..7008
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(7017..7102,1..22)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     7034..7050
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7084..7100)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
       61 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      121 acaacatagg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgaggtaac
      181 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      241 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
      301 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      361 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      421 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      481 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      541 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      601 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      661 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      721 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      781 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      841 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      901 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      961 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1021 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1081 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1141 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1201 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1261 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1321 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     1381 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1441 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1501 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1561 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1621 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1681 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1741 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1801 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1861 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1921 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1981 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2041 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     2101 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     2161 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     2221 cacctgacgt ctacaactca ttacgcacac tcatggtttt tggccagcga aaacagtttc
     2281 aaaagattgc tggaagtctg caataatgtc atcaataaat tcgataccaa cagagacacg
     2341 aattaagtcc ttggtaacac cagatgccaa cttttctttg tcatttaatt gtttgtgggt
     2401 agtgaagtat ggagcaatga ctaaggtctt ggcatcacca acattggcca agttagaggc
     2461 aagctttaaa ttgtcaacaa cttgagcacc agaaagtttg aatgggtcag tttccttgtc
     2521 ggcatttggt aagtctttta caccgaaaga taagacacca ccgaaaccgt tagatagata
     2581 cttcttagca ttttcatgat gagaatgaga tgctaaacca gggtatgaaa cccaagatac
     2641 gtatggggat tgttctaacc atttggctaa cttcaatgca ttttcaccgt gtctttcagc
     2701 tctcaaagat aatgtttcaa caccttgtag tagcaagaaa gaggcaaatg ggttcatcaa
     2761 tggacccaaa tctcttaata gttcagttct aacatgaacg atgtatgcca agttaccgta
     2821 ggcttcattg tagatagtac cgtgatatcc ttcggcaggt tgagagaatt gagggaactt
     2881 ttctgggtag tccttccatg ggaacttacc agagtcaaca ataataccac cgatagtagt
     2941 accatgacca ccaatccatt tggtagcaga atgtgttaca atatcagcac cgtatttaat
     3001 tggctgacag aagtaaccac cggcaccaaa tgtgttgtca acgacaactg gaataccgtg
     3061 tttgtgagca attgcaacaa ttttttcaaa atccggaaca ttgtactttg gattaccaat
     3121 ggtttccaaa taaacagcct tggttctttc atcaaagacc ttttcgaatt cttctggatt
     3181 gtcaccttca acaaatctag cctcgatacc aaatcttttg aacgagattt tgaactggtt
     3241 ataagtacca ccgtataagt aagaagtgga aacgatgttg tcaccagtgt gtgccaaacc
     3301 ttggatggca agggtttgag cggcttgacc ggaggaaaca gccaaagcag cagcaccacc
     3361 ttctaaagca gcaattcttt cttccaaaac attactggtt gggttttgga aacgggaata
     3421 gacgtaacct ggaacttcta gaccaaacaa ttgcgaacca tgcttagagt tttcgaaaac
     3481 ataagaagtg gtggcgtaaa ttggtacagc tctggatctg tgagcattgt caccagggtt
     3541 ctcttggccg gcgtgtagtt gaacagtatc gaaatgagat ggcattgtat ggatgggggt
     3601 aatagaattg tatctatgta tctgacgacc ctgtattaca cgaaggaaat agtagatgaa
     3661 aagacaagag agcaagaaaa aggaaaaact tcctttctaa cagacgcttt acttaacctt
     3721 atatatatct tatttttttc atcgagcgtg ttcaattgga caaggtgcca ccttttcgac
     3781 acttcggtta ttatgttaca cagttttcat gaggatggct atcatgacat taacctataa
     3841 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     3901 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     3961 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4021 ggcatcagag cagattgtac tgagagtgca ccatagatct gaattaattc ttgaataata
     4081 cataactttt cttaaaagaa tcaaagacag ataaaattta agagatatta aatattagtg
     4141 agaagccgag aattttgtaa caccaacata acactgacat ctttaacaac ttttaattat
     4201 gatacatttc ttacgtcatg attgattatt acagctatgc tgacaaatga ctcttgttgc
     4261 atggctacga accgggtaat actaagtgat tgactcttgc tgacctttta ttaagaacta
     4321 aatggacaat attatggagc atttcatgta taaattggtg cgtaaaatcg ttggatctct
     4381 cttctaagta catcctacta taacaatcaa gaaaaacaag aaaatcggac aaaacaatca
     4441 agtatggatt ctagaacagt tggtatatta ggagggggac aattgggacg tatgattgtt
     4501 gaggcagcaa acaggctcaa cattaagacg gtaatactag atgctgaaaa ttctcctgcc
     4561 aaacaaataa gcaactccaa tgaccacgtt aatggctcct tttccaatcc tcttgatatc
     4621 gaaaaactag ctgaaaaatg tgatgtgcta acgattgaga ttgagcatgt tgatgttcct
     4681 acactaaaga atcttcaagt aaaacatccc aaattaaaaa tttacccttc tccagaaaca
     4741 atcagattga tacaagacaa atatattcaa aaagagcatt taatcaaaaa tggtatagca
     4801 gttacccaaa gtgttcctgt ggaacaagcc agtgagacgt ccctattgaa tgttggaaga
     4861 gatttgggtt ttccattcgt cttgaagtcg aggactttgg catacgatgg aagaggtaac
     4921 ttcgttgtaa agaataagga aatgattccg gaagctttgg aagtactgaa ggatcgtcct
     4981 ttgtacgccg aaaaatgggc accatttact aaagaattag cagtcatgat tgtgaggtct
     5041 gttaacggtt tagtgttttc ttacccaatt gtagagacta tccacaagga caatatttgt
     5101 gacttatgtt atgcgcctgc tagagttccg gactccgttc aacttaaggc gaagttgttg
     5161 gcagaaaatg caatcaaatc ttttcccggt tgtggtatat ttggtgtgga aatgttctat
     5221 ttagaaacag gggaattgct tattaacgaa attgccccaa ggcctcacaa ctctggacat
     5281 tataccattg atgcttgcgt cacttctcaa tttgaagctc atttgagatc aatattggat
     5341 ttgccaatgc caaagaattt cacatctttc tccaccatta caacgaacgc cattatgcta
     5401 aatgttcttg gagacaaaca tacaaaagat aaagagctag aaacttgcga aagagcattg
     5461 gcgactccag gttcctcagt gtacttatat ggaaaagagt ctagacctaa cagaaaagta
     5521 ggtcacataa atattattgc ctccagtatg gcggaatgtg aacaaaggct gaactacatt
     5581 acaggtagaa ctgatattcc aatcaaaatc tctgtcgctc aaaagttgga cttggaagca
     5641 atggtcaaac cattggttgg aatcatcatg ggatcagact ctgacttgcc ggtaatgtct
     5701 gccgcatgtg cggttttaaa agattttggc gttccatttg aagtgacaat agtctctgct
     5761 catagaactc cacataggat gtcagcatat gctatttccg caagcaagcg tggaattaaa
     5821 acaattatcg ctggagctgg tggggctgct cacttgccag gtatggtggc tgcaatgaca
     5881 ccacttcctg tcatcggtgt gcccgtaaaa ggttcttgtc tagatggagt agattcttta
     5941 cattcaattg tgcaaatgcc tagaggtgtt ccagtagcta ccgtcgctat taataatagt
     6001 acgaacgctg cgctgttggc tgtcagactg cttggcgctt atgattcaag ttatacaacg
     6061 aaaatggaac agtttttatt aaagcaagaa gaagaagttc ttgtcaaagc acaaaagtta
     6121 gaaactgtcg gttacgaagc ttatctagaa aacaagtaat atataagttt attgatatac
     6181 ttgtacagca aataattata aaatgatata cctatttttt aggctttgtt atgattacat
     6241 caaatgtgga cttcatacat agaaatcaac gcttacaggt gtcctttttt aagaatttca
     6301 tacataagat ctatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6361 aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6421 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     6481 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     6541 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6601 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6661 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6721 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6781 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg ccattcaggc
     6841 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     6901 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     6961 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
     7021 ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc ctgcagcccg
     7081 ggggatccac tagttctaga gc
//
