LOCUS       Exported                7978 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 7978)
  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..7978
                     /organism="synthetic DNA construct"
                     /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..2789
                     /label=oRH4982
     promoter        2902..3306
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3319..4413
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVAGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(4659..4708)
                     /label=oRH4983
     promoter        4926..5301
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             5302..7017
                     /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(7222..7677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     7822..7838
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7841..7867
                     /label=Recomb OH 2
     promoter        7848..7866
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7892..7908
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7942..7958)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(7958..7978,1..9))
                     /label=Recomb OH
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 acgtctcctg tacttccttg ttcatgtgtg ttcaaaaacg ttatatttat aggataatta
     2821 tactctattt ctcaacaagt aattggttgt ttggccgagc ggtctaaggc gcctgattca
     2881 agaaatatct tgaccgcagt taactgtggg aatactcagg tatcgtaaga tgcaagagtt
     2941 cgaatctctt agcaaccatt atttttttcc tcaacataac gagaacacac aggggcgcta
     3001 tcgcacagaa tcaaattcga tgactggaaa ttttttgtta atttcagagg tcgcctgacg
     3061 catatacctt tttcaactga aaaattggga gaaaaaggaa aggtgagagg ccggaaccgg
     3121 cttttcatat agaatagaga agcgttcatg actaaatgct tgcatcacaa tacttgaagt
     3181 tgacaatatt atttaaggac ctattgtttt ttccaatagg tggttagcaa tcgtcttact
     3241 ttctaacttt tcttaccttt tacatttcag caatatatat atatatttca aggatatacc
     3301 attctaatgt ctgcccctat gtctgcccct aagaagatcg tcgttttgcc aggtgaccac
     3361 gttggtcaag aaatcacagc cgaagccatt aaggttctta aagctatttc tgatgttcgt
     3421 tccaatgtca agttcgattt cgaaaatcat ttaattggtg gtgctgctat cgatgctaca
     3481 ggtgtcccac ttccagatga ggcgctggaa gcctccaaga aggttgatgc cgttttgtta
     3541 ggtgctgtgg ctggtcctaa atggggtacc ggtagtgtta gacctgaaca aggtttacta
     3601 aaaatccgta aagaacttca attgtacgcc aacttaagac catgtaactt tgcatccgac
     3661 tctcttttag acttatctcc aatcaagcca caatttgcta aaggtactga cttcgttgtt
     3721 gtcagagaat tagtgggagg tatttacttt ggtaagagaa aggaagacga tggtgatggt
     3781 gtcgcttggg atagtgaaca atacaccgtt ccagaagtgc aaagaatcac aagaatggcc
     3841 gctttcatgg ccctacaaca tgagccacca ttgcctattt ggtccttgga taaagctaat
     3901 cttttggcct cttcaagatt atggagaaaa actgtggagg aaaccatcaa gaacgaattc
     3961 cctacattga aggttcaaca tcaattgatt gattctgccg ccatgatcct agttaagaac
     4021 ccaacccacc taaatggtat tataatcacc agcaacatgt ttggtgatat catctccgat
     4081 gaagcctccg ttatcccagg ttccttgggt ttgttgccat ctgcgtcctt ggcctctttg
     4141 ccagacaaga acaccgcatt tggtttgtac gaaccatgcc acggttctgc tccagatttg
     4201 ccaaagaata aggttgaccc tatcgccact atcttgtctg ctgcaatgat gttgaaattg
     4261 tcattgaact tgcctgaaga aggtaaggcc attgaagatg cagttaaaaa ggttttggat
     4321 gcaggtatca gaactggtga tttaggtggt tccaacagta ccaccgaagt cggtgatgct
     4381 gtcgccgaag aagttaagaa aatccttgct taaaaagatt ctcttttttt atgatatttg
     4441 tacataaact ttataaatga aattcataat agaaacgaca cgaaattaca aaatggaata
     4501 tgttcatagg gtagacgaaa ctatatacgc aatctacata catttatcaa gaaggagaaa
     4561 aaggaggata gtaaaggaat acaggtaagc aaattgatac taatggctca acgtgataag
     4621 gaaaaagaat tgcactttaa cattaatatt gacaaggagg agggcaccac acaaaaagat
     4681 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4741 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4801 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4861 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atagatctga
     4921 attaattctt gaataataca taacttttct taaaagaatc aaagacagat aaaatttaag
     4981 agatattaaa tattagtgag aagccgagaa ttttgtaaca ccaacataac actgacatct
     5041 ttaacaactt ttaattatga tacatttctt acgtcatgat tgattattac agctatgctg
     5101 acaaatgact cttgttgcat ggctacgaac cgggtaatac taagtgattg actcttgctg
     5161 accttttatt aagaactaaa tggacaatat tatggagcat ttcatgtata aattggtgcg
     5221 taaaatcgtt ggatctctct tctaagtaca tcctactata acaatcaaga aaaacaagaa
     5281 aatcggacaa aacaatcaag tatggattct agaacagttg gtatattagg agggggacaa
     5341 ttgggacgta tgattgttga ggcagcaaac aggctcaaca ttaagacggt aatactagat
     5401 gctgaaaatt ctcctgccaa acaaataagc aactccaatg accacgttaa tggctccttt
     5461 tccaatcctc ttgatatcga aaaactagct gaaaaatgtg atgtgctaac gattgagatt
     5521 gagcatgttg atgttcctac actaaagaat cttcaagtaa aacatcccaa attaaaaatt
     5581 tacccttctc cagaaacaat cagattgata caagacaaat atattcaaaa agagcattta
     5641 atcaaaaatg gtatagcagt tacccaaagt gttcctgtgg aacaagccag tgagacgtcc
     5701 ctattgaatg ttggaagaga tttgggtttt ccattcgtct tgaagtcgag gactttggca
     5761 tacgatggaa gaggtaactt cgttgtaaag aataaggaaa tgattccgga agctttggaa
     5821 gtactgaagg atcgtccttt gtacgccgaa aaatgggcac catttactaa agaattagca
     5881 gtcatgattg tgaggtctgt taacggttta gtgttttctt acccaattgt agagactatc
     5941 cacaaggaca atatttgtga cttatgttat gcgcctgcta gagttccgga ctccgttcaa
     6001 cttaaggcga agttgttggc agaaaatgca atcaaatctt ttcccggttg tggtatattt
     6061 ggtgtggaaa tgttctattt agaaacaggg gaattgctta ttaacgaaat tgccccaagg
     6121 cctcacaact ctggacatta taccattgat gcttgcgtca cttctcaatt tgaagctcat
     6181 ttgagatcaa tattggattt gccaatgcca aagaatttca catctttctc caccattaca
     6241 acgaacgcca ttatgctaaa tgttcttgga gacaaacata caaaagataa agagctagaa
     6301 acttgcgaaa gagcattggc gactccaggt tcctcagtgt acttatatgg aaaagagtct
     6361 agacctaaca gaaaagtagg tcacataaat attattgcct ccagtatggc ggaatgtgaa
     6421 caaaggctga actacattac aggtagaact gatattccaa tcaaaatctc tgtcgctcaa
     6481 aagttggact tggaagcaat ggtcaaacca ttggttggaa tcatcatggg atcagactct
     6541 gacttgccgg taatgtctgc cgcatgtgcg gttttaaaag attttggcgt tccatttgaa
     6601 gtgacaatag tctctgctca tagaactcca cataggatgt cagcatatgc tatttccgca
     6661 agcaagcgtg gaattaaaac aattatcgct ggagctggtg gggctgctca cttgccaggt
     6721 atggtggctg caatgacacc acttcctgtc atcggtgtgc ccgtaaaagg ttcttgtcta
     6781 gatggagtag attctttaca ttcaattgtg caaatgccta gaggtgttcc agtagctacc
     6841 gtcgctatta ataatagtac gaacgctgcg ctgttggctg tcagactgct tggcgcttat
     6901 gattcaagtt atacaacgaa aatggaacag tttttattaa agcaagaaga agaagttctt
     6961 gtcaaagcac aaaagttaga aactgtcggt tacgaagctt atctagaaaa caagtaatat
     7021 ataagtttat tgatatactt gtacagcaaa taattataaa atgatatacc tattttttag
     7081 gctttgttat gattacatca aatgtggact tcatacatag aaatcaacgc ttacaggtgt
     7141 ccttttttaa gaatttcata cataagatct atgcggtgtg aaataccgca cagatgcgta
     7201 aggagaaaat accgcatcag gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa
     7261 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     7321 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     7381 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     7441 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     7501 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     7561 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     7621 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcgc
     7681 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     7741 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     7801 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     7861 tatagggcga attgggtacc gggccccccc tcgaggtcga cggtatcgat aagcttgata
     7921 tcgaattcct gcagcccggg ggatccacta gttctagagc ggccgccctc ttagcgct
//
