LOCUS       Exported                7112 bp ds-DNA     circular SYN 29-MAR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7112)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2019 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7112
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    11..514
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        complement(569..587)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(608..624)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    632..648
                     /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(656..686)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(1010..1598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1769..2629)
                     /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(2630..2734)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2737..2786
                     /label=oRH5046
     promoter        2816..3002
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3003..3662
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     primer_bind     complement(3791..3839)
                     /label=oRH5045
     promoter        4060..4435
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             4436..6151
                     /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(6356..6811)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6956..6972
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6975..7001
                     /label=Recomb OH 1
     promoter        6982..7000
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7026..7042
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7076..7092)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(7092..7112,1..9))
                     /label=Recomb OH 2
ORIGIN
        1 tcttaggacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat
       61 aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag
      121 taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa
      181 aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat
      241 agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc
      301 ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat
      361 aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac
      421 tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa
      481 aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg gcgcggccgc
      541 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
      601 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      661 taggagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagg taactcacat
      721 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      781 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      841 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1441 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1561 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1621 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1681 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1741 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1801 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1861 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1921 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1981 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2041 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2101 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2161 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2221 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2281 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2341 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2401 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2461 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2521 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2581 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2641 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2701 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2761 acgtctaaca cagtcctttc ccgcaatttt ctttttctat tactcttggc ctcctctagt
     2821 acactctata tttttttatg cctcggtaat gattttcatt tttttttttc ccctagcgga
     2881 tgactctttt tttttcttag cgattggcat tatcacataa tgaattatac attatataaa
     2941 gtaatgtgat ttcttcgaag aatatactaa aaaatgagca ggcaagataa acgaaggcaa
     3001 agatgacaga gcagaaagcc ctagtaaagc gtattacaaa tgaaaccaag attcagattg
     3061 cgatctcttt aaagggtggt cccctagcga tagagcactc gatcttccca gaaaaagagg
     3121 cagaagcagt agcagaacag gccacacaat cgcaagtgat taacgtccac acaggtatag
     3181 ggtttctgga ccatatgata catgctctgg ccaagcattc cggctggtcg ctaatcgttg
     3241 agtgcattgg tgacttacac atagacgacc atcacaccac tgaagactgc gggattgctc
     3301 tcggtcaagc ttttaaagag gccctactgg cgcgtggagt aaaaaggttt ggatcaggat
     3361 ttgcgccttt ggatgaggca ctttccagag cggtggtaga tctttcgaac aggccgtacg
     3421 cagttgtcga acttggtttg caaagggaga aagtaggaga tctctcttgc gagatgatcc
     3481 cgcattttct tgaaagcttt gcagaggcta gcagaattac cctccacgtt gattgtctgc
     3541 gaggcaagaa tgatcatcac cgtagtgaga gtgcgttcaa ggctcttgcg gttgccataa
     3601 gagaagccac ctcgcccaat ggtaccaacg atgttccctc caccaaaggt gttcttatgt
     3661 agtgacaccg attatttaaa gctgcagcat acgatatata tacatgtgta tatatgtata
     3721 cctatgaatg tcagtaagta tgtatacgaa cagtatgata ctgaagatga caaggtaatg
     3781 catcattcta tacgtgtcat tctgaacgag gcatcatgac attaacctat aaaaataggc
     3841 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3901 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3961 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     4021 agcagattgt actgagagtg caccatagat ctgaattaat tcttgaataa tacataactt
     4081 ttcttaaaag aatcaaagac agataaaatt taagagatat taaatattag tgagaagccg
     4141 agaattttgt aacaccaaca taacactgac atctttaaca acttttaatt atgatacatt
     4201 tcttacgtca tgattgatta ttacagctat gctgacaaat gactcttgtt gcatggctac
     4261 gaaccgggta atactaagtg attgactctt gctgaccttt tattaagaac taaatggaca
     4321 atattatgga gcatttcatg tataaattgg tgcgtaaaat cgttggatct ctcttctaag
     4381 tacatcctac tataacaatc aagaaaaaca agaaaatcgg acaaaacaat caagtatgga
     4441 ttctagaaca gttggtatat taggaggggg acaattggga cgtatgattg ttgaggcagc
     4501 aaacaggctc aacattaaga cggtaatact agatgctgaa aattctcctg ccaaacaaat
     4561 aagcaactcc aatgaccacg ttaatggctc cttttccaat cctcttgata tcgaaaaact
     4621 agctgaaaaa tgtgatgtgc taacgattga gattgagcat gttgatgttc ctacactaaa
     4681 gaatcttcaa gtaaaacatc ccaaattaaa aatttaccct tctccagaaa caatcagatt
     4741 gatacaagac aaatatattc aaaaagagca tttaatcaaa aatggtatag cagttaccca
     4801 aagtgttcct gtggaacaag ccagtgagac gtccctattg aatgttggaa gagatttggg
     4861 ttttccattc gtcttgaagt cgaggacttt ggcatacgat ggaagaggta acttcgttgt
     4921 aaagaataag gaaatgattc cggaagcttt ggaagtactg aaggatcgtc ctttgtacgc
     4981 cgaaaaatgg gcaccattta ctaaagaatt agcagtcatg attgtgaggt ctgttaacgg
     5041 tttagtgttt tcttacccaa ttgtagagac tatccacaag gacaatattt gtgacttatg
     5101 ttatgcgcct gctagagttc cggactccgt tcaacttaag gcgaagttgt tggcagaaaa
     5161 tgcaatcaaa tcttttcccg gttgtggtat atttggtgtg gaaatgttct atttagaaac
     5221 aggggaattg cttattaacg aaattgcccc aaggcctcac aactctggac attataccat
     5281 tgatgcttgc gtcacttctc aatttgaagc tcatttgaga tcaatattgg atttgccaat
     5341 gccaaagaat ttcacatctt tctccaccat tacaacgaac gccattatgc taaatgttct
     5401 tggagacaaa catacaaaag ataaagagct agaaacttgc gaaagagcat tggcgactcc
     5461 aggttcctca gtgtacttat atggaaaaga gtctagacct aacagaaaag taggtcacat
     5521 aaatattatt gcctccagta tggcggaatg tgaacaaagg ctgaactaca ttacaggtag
     5581 aactgatatt ccaatcaaaa tctctgtcgc tcaaaagttg gacttggaag caatggtcaa
     5641 accattggtt ggaatcatca tgggatcaga ctctgacttg ccggtaatgt ctgccgcatg
     5701 tgcggtttta aaagattttg gcgttccatt tgaagtgaca atagtctctg ctcatagaac
     5761 tccacatagg atgtcagcat atgctatttc cgcaagcaag cgtggaatta aaacaattat
     5821 cgctggagct ggtggggctg ctcacttgcc aggtatggtg gctgcaatga caccacttcc
     5881 tgtcatcggt gtgcccgtaa aaggttcttg tctagatgga gtagattctt tacattcaat
     5941 tgtgcaaatg cctagaggtg ttccagtagc taccgtcgct attaataata gtacgaacgc
     6001 tgcgctgttg gctgtcagac tgcttggcgc ttatgattca agttatacaa cgaaaatgga
     6061 acagttttta ttaaagcaag aagaagaagt tcttgtcaaa gcacaaaagt tagaaactgt
     6121 cggttacgaa gcttatctag aaaacaagta atatataagt ttattgatat acttgtacag
     6181 caaataatta taaaatgata tacctatttt ttaggctttg ttatgattac atcaaatgtg
     6241 gacttcatac atagaaatca acgcttacag gtgtcctttt ttaagaattt catacataag
     6301 atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     6361 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     6421 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     6481 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     6541 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     6601 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6661 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6721 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6781 gccgcgctta atgcgccgct acagggcgcg tcgcgccatt cgccattcag gctgcgcaac
     6841 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     6901 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     6961 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc
     7021 cccctcgagg tcgacggtat cgataagctt gatatcgaat tcctgcagcc cgggggatcc
     7081 actagttcta gagcggccgc cctcttagcg ct
//
