LOCUS       Exported                5382 bp ds-DNA     circular SYN 31-JAN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5382)
  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..5382
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     7..23
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(57..73)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(98..124)
                     /label=Recomb OH 1
     promoter        complement(99..117)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(127..143)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      288..743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(874..1677)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1678..1893)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     2103..2162
                     /label=oRH4869
     CDS             complement(2191..2865)
                     /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"
     promoter        complement(2866..3146)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3135..3194)
                     /label=oRH4868
     promoter        3187..3291
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3292..4152
                     /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"
     rep_origin      4323..4911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5235..5265
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5273..5289
                     /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)."
     primer_bind     5297..5313
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5334..5352
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     join(5380..5382,1..7)
                     /label=Recomb OH 2
ORIGIN
        1 ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa gcttatcgat
       61 accgtcgacc tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc
      121 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      181 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      241 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag
      301 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
      361 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
      421 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
      481 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
      541 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
      601 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
      661 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
      721 caaaatatta acgtttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
      781 atttcacacc gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt
      841 tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa
      901 atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt
      961 tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca catcatccac
     1021 ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat
     1081 aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat
     1141 aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga
     1201 tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac
     1261 ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt
     1321 cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt
     1381 attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa
     1441 tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt
     1501 ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt
     1561 acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt
     1621 ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat
     1681 ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt
     1741 tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct
     1801 tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaagaa accgaaatca
     1861 aaaaaaagaa taaaaaaaaa atgatgaatt gaattgaaaa gctgtggtat ggtgcactct
     1921 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     1981 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2041 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2101 gggcctcgtg atacgcctat ttttataggt taatgtcatg attaggcaag tgcacaaaca
     2161 atacttaaat aaatactact cagtaataac ctatttctta gcatttttga cgaaatttgc
     2221 tattttgtta gagtctttta caccatttgt ctccacacct ccgcttacat caacaccaat
     2281 aacgccattt aatctaagcg catcaccaac attttctggc gtcagtccac cagctaacat
     2341 aaaatgtaag ctttcggggc tctcttgcct tccaacccag tcagaaatcg agttccaatc
     2401 caaaagttca cctgtcccac ctgcttctga atcaaacaag ggaataaacg aatgaggttt
     2461 ctgtgaagct gcactgagta gtatgttgca gtcttttgga aatacgagtc ttttaataac
     2521 tggcaaaccg aggaactctt ggtattcttg ccacgactca tctccatgca gttggacgat
     2581 atcaatgccg taatcattga ccagagccaa aacatcctcc ttaggttgat tacgaaacac
     2641 gccaaccaag tatttcggag tgcctgaact atttttatat gcttttacaa gacttgaaat
     2701 tttccttgca ataaccgggt caattgttct ctttctattg ggcacacata taatacccag
     2761 caagtcagca tcggaatcta gagcacattc tgcggcctct gtgctctgca agccgcaaac
     2821 tttcaccaat ggaccagaac tacctgtgaa attaataaca gacatactcc aagctgcctt
     2881 tgtgtgctta atcacgtata ctcacgtgct caatagtcac caatgccctc cctcttggcc
     2941 ctctcctttt cttttttcga ccgaattaat tcttaatcgg caaaaaaaga aaagctccgg
     3001 atcaagattg tacgtaaggt gacaagctat ttttcaataa agaatatctt ccactactgc
     3061 catctggcgt cataactgca aagtacacat atattacgat gctgtctatt aaatgcttcc
     3121 tatattatat atatagtaat gtcgtttatg gtgcagacgt caggtggcac ttttcgggga
     3181 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3241 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3301 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3361 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3421 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3481 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3541 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3601 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3661 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3721 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3781 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3841 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3901 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3961 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4021 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4081 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4141 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4201 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4261 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4321 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4381 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4441 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4501 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4561 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4621 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4681 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4741 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4801 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4861 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4921 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4981 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5041 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     5101 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5161 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta cctcactcat
     5221 taggcacccc aggctttaca ctttatgctt ccggctccta tgttgtgtgg aattgtgagc
     5281 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     5341 cctcactaaa gggaacaaaa gctggagctc caccgcggtg gc
//
