LOCUS       Exported                5934 bp ds-DNA     circular SYN 12-JAN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5934)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2019 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5934
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     21..37
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(71..87)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(112..138)
                     /label=Recomb OH 1
     promoter        complement(113..131)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(141..157)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      302..757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(888..1691)
                     /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(1692..1907)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     2117..2176
                     /label=oRH4869
     CDS             complement(2205..2879)
                     /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(2880..3160)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3149..3208)
                     /label=oRH4868
     promoter        3201..3305
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3306..4166
                     /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      4337..4925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5249..5279
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5287..5303
                     /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     5311..5327
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5348..5366
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    5421..5924
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     join(5926..5934,1..21)
                     /label=Recomb OH 2
ORIGIN
        1 agcgctaaga gggcggccgc tctagaacta gtggatcccc cgggctgcag gaattcgata
       61 tcaagcttat cgataccgtc gacctcgagg gggggcccgg tacccaattc gccctatagt
      121 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
      181 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
      241 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
      301 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      361 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      421 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      481 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      541 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      601 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      661 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      721 aacgcgaatt ttaacaaaat attaacgttt acaatttcct gatgcggtat tttctcctta
      781 cgcatctgtg cggtatttca caccgcatag ggtaataact gatataatta aattgaagct
      841 ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta gttttgctgg
      901 ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta
      961 gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag
     1021 accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc
     1081 acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga
     1141 gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta
     1201 tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta
     1261 ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc
     1321 acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac
     1381 tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg
     1441 tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag
     1501 atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt
     1561 aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg
     1621 atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta
     1681 gctttcgaca tgatttatct tcgtttcctg caggtttttg ttctgtgcag ttgggttaag
     1741 aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt
     1801 ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa
     1861 agaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaattg aaaagctgtg
     1921 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     1981 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     2041 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     2101 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgattagg
     2161 caagtgcaca aacaatactt aaataaatac tactcagtaa taacctattt cttagcattt
     2221 ttgacgaaat ttgctatttt gttagagtct tttacaccat ttgtctccac acctccgctt
     2281 acatcaacac caataacgcc atttaatcta agcgcatcac caacattttc tggcgtcagt
     2341 ccaccagcta acataaaatg taagctttcg gggctctctt gccttccaac ccagtcagaa
     2401 atcgagttcc aatccaaaag ttcacctgtc ccacctgctt ctgaatcaaa caagggaata
     2461 aacgaatgag gtttctgtga agctgcactg agtagtatgt tgcagtcttt tggaaatacg
     2521 agtcttttaa taactggcaa accgaggaac tcttggtatt cttgccacga ctcatctcca
     2581 tgcagttgga cgatatcaat gccgtaatca ttgaccagag ccaaaacatc ctccttaggt
     2641 tgattacgaa acacgccaac caagtatttc ggagtgcctg aactattttt atatgctttt
     2701 acaagacttg aaattttcct tgcaataacc gggtcaattg ttctctttct attgggcaca
     2761 catataatac ccagcaagtc agcatcggaa tctagagcac attctgcggc ctctgtgctc
     2821 tgcaagccgc aaactttcac caatggacca gaactacctg tgaaattaat aacagacata
     2881 ctccaagctg cctttgtgtg cttaatcacg tatactcacg tgctcaatag tcaccaatgc
     2941 cctccctctt ggccctctcc ttttcttttt tcgaccgaat taattcttaa tcggcaaaaa
     3001 aagaaaagct ccggatcaag attgtacgta aggtgacaag ctatttttca ataaagaata
     3061 tcttccacta ctgccatctg gcgtcataac tgcaaagtac acatatatta cgatgctgtc
     3121 tattaaatgc ttcctatatt atatatatag taatgtcgtt tatggtgcag acgtcaggtg
     3181 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3241 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3301 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3361 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3421 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3481 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3541 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3601 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3661 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3721 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3781 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3841 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3901 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3961 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4021 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4081 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4141 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4201 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4261 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4321 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4381 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4441 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4501 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4561 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4621 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4681 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4741 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4801 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4861 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4921 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4981 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5041 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5101 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5161 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     5221 gttacctcac tcattaggca ccccaggctt tacactttat gcttccggct cctatgttgt
     5281 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     5341 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc
     5401 gcgccacctg ggtccttttc atcacgtgct ataaaaataa ttataattta aattttttaa
     5461 tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt
     5521 gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc ttttatcttg
     5581 ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt agaagaccac
     5641 acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat atctatttaa
     5701 tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga aattttttaa
     5761 acctttgttt attttttttt cttcattccg taactcttct accttcttta tttactttct
     5821 aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca aattattcca
     5881 tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc gatccgtcct aaga
//
