LOCUS       Exported                4936 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 4936)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2019 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4936
                     /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
     promoter        2203..2307
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2308..3168
                     /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      3339..3927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4251..4281
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4289..4305
                     /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     4313..4329
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4350..4368
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     4397..4424
                     /label=oRH4866
     misc_feature    4423..4926
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(join(4917..4936,1..20))
                     /label=oRH4867
     primer_bind     join(4928..4936,1..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 catgataata
     2161 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2221 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2281 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2341 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2401 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2461 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2521 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2581 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2641 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2701 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2761 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2821 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     2881 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2941 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3001 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3061 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3121 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3181 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3241 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3301 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3361 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3421 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3481 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3541 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3601 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3661 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3721 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3781 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3841 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3901 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3961 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4021 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4081 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4141 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     4201 gagcgcaacg caattaatgt gagttacctc actcattagg caccccaggc tttacacttt
     4261 atgcttccgg ctcctatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     4321 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     4381 gagctccacc gcggtggcgg ccgcgccacc tgggtccttt tcatcacgtg ctataaaaat
     4441 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
     4501 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
     4561 acaaatacta ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca
     4621 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
     4681 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
     4741 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
     4801 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
     4861 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
     4921 gcgatccgtc ctaaga
//
