LOCUS       Exported                8285 bp ds-DNA     circular SYN 16-MAY-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    SnapGene DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8285)
  AUTHORS   Boeke Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 19, 2017 from SnapGene 3.3.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8285
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             217..795
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISRLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    273..380
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(389..407)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(417..433)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      578..1033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    complement(1158..2492)
                     /label=S. pombe leu1
     CDS             complement(1256..2371)
                     /codon_start=1
                     /label=leu1 CDS
                     /translation="MCAKKIVVLPGDHIGPEIVASALEVLKVVEKKRPELKLEFEEHKI
                     GGASIDAYGTPLTDETVKACLEADGVLLGAVGGPEWTNPNCRPEQGLLKLRKSMGVWAN
                     LRPCNFASKSLVKYSPLKPEIVEGVDFCVVRELTGGCYFGERTEDNGSGYAMDTWPYSL
                     EEVSRIARLAAWLAETSNPPAPVTLLDKANVLATSRLWRKTVAKIFKEEYPHLTLKNQL
                     IDSAAMLLVKSPRTLNGVVLTDNLFGDIISDEASVIPGSLGLLPSASLSGVVGKSEEKV
                     HCLVEPIHGSAPDIAGKGIVNPVGTILSASLLLRYGLNAPKEAEAIEAAVRKVLDDTSI
                     GGRGLYTRDLGGEASTADITKAVVEELEKIL"
     promoter        2959..4123
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     terminator      4150..5141
                     /label=nmt1 terminator
     rep_origin      5221..6004
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6380..6484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6485..7345
                     /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      7516..8104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgctcta
      301 gaactagtcg atccgccggg ctgcaggaat tcgatatcaa ggttatcgat accgtccacc
      361 tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
      421 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
      481 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
      541 tcccaacagt tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta
      601 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
      661 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      721 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      781 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      841 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
      901 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
      961 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     1021 acgtttacaa tttcaactcg gtcataaagt tgaacggatg tcgtaaatca attccatgct
     1081 tttgcaactt aatacatctt tgcttgacta taacatacca aaagaaaaaa cattagagcc
     1141 ttaaaattta cgcttttgtc aattgcattt ttaacgataa taaatatgcg acacaattaa
     1201 tcaataaaaa ttaggtcaaa tcaaatgata tgaatttatt ctacattaaa ccctattaca
     1261 aaattttttc aagttcttca acaacagcct tagtaatatc agcggtagaa gcctcacctc
     1321 ccaaatcgcg agtataaaga ccacgtccac caattgaagt atcatccaag accttgcgta
     1381 cggcggcttc gatagcttca gcctccttag gagcattcaa accataacga aggagaaggg
     1441 aagcagataa aattgtacca acaggattaa caatgccctt gccagcgata tcgggagcgc
     1501 taccgtgaat gggctcaacc aaacaatgaa ccttttcttc tgattttcct accacaccgg
     1561 aaagggaggc agaaggcaaa aggcccaagc taccaggaat gacagaagcc tcatctgaaa
     1621 taatgtcacc aaacaagttg tcagtcaaaa caacaccgtt aagtgtacga gggctcttga
     1681 ccaaaagcat ggctgcggag tcaatgagct ggttttttaa ggtaaggtga ggatattcct
     1741 ccttaaaaat cttagctaca gtcttgcgcc aaagacgaga agttgccaaa acattagctt
     1801 tgtcgagtaa tgtgacggga gcaggagggt tggaagtttc agctaaccaa gcagccaaac
     1861 gagcaatacg agaaacttct tccaaactgt aaggccaagt gtccatagca taacccgatc
     1921 cgttgtcctc agtgcgctca ccaaagtaac aacctccagt aagttctcgt acaacacaaa
     1981 aatcgacacc ttcaacgatt tcaggcttca aagggctgta cttgactaaa gacttgctgg
     2041 caaagttgca aggtcgaagg ttggcccaaa cacccatact cttacgaagc ttcaataaac
     2101 cttgctcagg acgacaattg gggttggtcc attcaggacc accaacggca cccaaaagaa
     2161 caccgtcagc ttccaaacaa gccttcacag tctcgtcagt caaaggggtt ccataggcat
     2221 caatagaggc acctccaatc ttgtgttctt caaactccag ttttaactca ggtcgcttct
     2281 tctcaacgac tttcaaaacc tccaaggcag aagcaacaat ttcagggcca atatgatctc
     2341 ctggtaagac gacgattttc tttgcacaca tgttgttgaa gaagttttgt tgtgaaatgg
     2401 tttcgtgaaa gtttcagacc ctaccgcaaa aatgcctggt ttcgggaaac tcaacactgt
     2461 tgcacttttt atactacaga ttgggatatc gataatattg cgtaaaaaat ccttttttta
     2521 aaaagcttgt ttacagtaac gtaaatgacc agaaatcaga tgaaaatcac aagaaagcaa
     2581 ataattcacg ttaaatcctg atatgtttga ttttgtgatg aaatcatgga tgttcatagg
     2641 aattgttgaa attgcgcttt tttaacgaaa tatacaagta tcctggagct tacttaatta
     2701 attaatgaat ctttgtttct agatattaaa atagtagcct caattatcag cgctttctac
     2761 gttagtaaac gaaattttta atgtcaaaaa aatgtttaat agacagtacg aatatgcatt
     2821 ataatgttca taaataattt gagtatgtgt acgagttgtc tttaaaccca cagaggtaga
     2881 atgtatatat aaaattaata agctaagtgt aatacttaaa aaatacatgt cgatcgactc
     2941 tagaggatca gaaaattatc gccataaaag acagaataag tcatcagcgg ttgtttcatt
     3001 tcctatattt tttttttatt tttttatttt ttaataaggg aaaatttaac gtctaaggat
     3061 acagaagatt gttagcacat taaagtaata aaggcttaag tagtaagtgc cttagcatgt
     3121 tattgtattt caaaggacat aatctaaaat aataacaata tcatttctca caagttattc
     3181 aattttcttt tttttttcta ataatatcaa gaatgtatta tttgtttgac ataagtcaac
     3241 taatttattt aatatgctgg attaatcttg cagacatgta aattaacaag ttttagtcaa
     3301 ataacgttga agtttcaatg aactcaaata atttctcttt ttttttatat aaccataagt
     3361 ctaatctgat ttatattttc cgcagggatc aactgaagtt atgacatttg gattggatca
     3421 cttataacct tggtcgccaa ataatacaaa aatcagcgtt ataaaacaaa gaaggttttt
     3481 gttaagaaat taatcctctt tcttgataag aaagttgaac cgaaattgca gatactgata
     3541 tatgaaaata atacccacaa ttttgggaat agcgcaagcc tcaatttaaa caataggtga
     3601 ggacacatga taatgacctc aatgattgtt agaagaaaag agcctcatta caaaatcgaa
     3661 aaatgaatgg ttgggtacaa gtttccaaaa catggtaaag tggactttgc gtatgagacg
     3721 taaatagaaa aaaacacttg ttatatgttt tctagaatta ttgttgtctc tttatggttg
     3781 gatgatgcaa aatagtaatt tcggttagtt gctgtaaaac accacgagac aaatagatat
     3841 ggatatttat taaatcagga aaaacgtaac tctcggctac tggatggttc agtcacccaa
     3901 cgattactgg ggagagaaaa cagggcaaaa gcaaagctta aaggaatccg attgtcattc
     3961 ggcaatgtgc agcgaaacta aaaaccggat aatggacctg ttaatcgaaa cattgaagat
     4021 atataaagga agaggaatcc tggcatatca tcaattgaat aagttgaatt aattatttca
     4081 atctcattct cactttctga cttatagtcg ctttgttaaa tcatatgtcg actctagagg
     4141 atccccgggt aaaaggaatg tctcccttgc cagtactgct agggtttttc tttcaaacta
     4201 tggaagccca ttcaagctgc atattacgat tttgtttttc gcttttagaa agtggtttag
     4261 atgagataat agaaaaattc ttgatctccg acaacgagta cttttatttt ttttgctaat
     4321 cactttactc aatattagct cgaaatcgta gaaacgtaga cgggtgcggg ataccgagtg
     4381 gtgtagttaa gaatttttat aaaccacgtg gcccaaaaat atgaacccaa aacgtttata
     4441 catgagtata ctttaagaag gctatacccc ttcgtgttag atgtagtttt agctacccaa
     4501 cccgagtcta tgagcttgac ttcagatgta gaaggcatta aatcgttttg aatattaatt
     4561 aaaaaacgat gaaaattaaa tatttaaaag caatcatacg ctgaaaattt agtgctgtgg
     4621 ctaatccttc aacatggaaa tgccataaaa gtgactttga caaaaaaaaa agtatataca
     4681 ggtagtaaac tcatctactt cattgacttt gtttacagca tgtggaagga ggaatattta
     4741 ttgctaaatc gtagtttaac attcaataag taatactatt gaaattcgac aagattggcc
     4801 gcatggatga aaaagaggca ttttgctttg ggagaattag ttcaaattag aactgaaaaa
     4861 aaaaacttta cgaggcaaaa atgtcggatt gagatcgtaa aagttcgctc gtcgtctttt
     4921 gctttgtgat tgttttcatg gatacatctt gctggatatt taaattttag tactatgtat
     4981 aagatattct ataaatgttt tatcacccaa acctgttagc gccttcttaa ttctattcaa
     5041 tctggctttt gctctgagac tacttcttgg actttcacta cttgttagtt atacggaatt
     5101 tgtgtaatta gaagtgaaat aatcctttct attagtaatg cgagctgcaa ttcgagtcta
     5161 actccttaac cactcggaca tagtgactta tctgacactc attgtaaatt aatatatata
     5221 taggcatttt gtttagttaa aggtacttaa gtaattagta taaacgaacc aattttataa
     5281 tcaggaagtt aagtgaatgg tagcacatgt cgtaaaaatt gtgaattttt attgaataat
     5341 attttaaata caagcctttc tagactaggt atactcataa acatatatga gcaaaaggat
     5401 agaggagatt acattgcatc ttctacaaat ttatttattg ccctttactg aaaaattaaa
     5461 taatgagtac taaatgataa aaagcgctca gtacaagaaa gattgcaaaa atattgcatt
     5521 cttcatgaat taaagttgca tataaggcat attgaaagta atagtactaa aacagcagtt
     5581 agcgaaaatt aatagaatta tattcgcaag acaattgtga cattaaatta aaaaattgta
     5641 gaatttttac tatcctcttt aacgccatga gcctttataa aaaggttaaa ttagttttaa
     5701 cattcttttt ttgagtaaga gtttacaatt tatcaaaacc tgtgttatta tattcattag
     5761 tttcaattta ttagcatcta gagaaaaatc aattggcagt tacattgttg gaatttatga
     5821 agaaaaagaa tctacaacgg agaataagtt gctgatcgct ttccctaaaa ttgtatattt
     5881 tgctgagctt attttgacat ttcgttgaag ttttctctaa ttcgcattca ttttaagtaa
     5941 aacaatgaga aataaaatta caaaaaatac aaattaaaat acaattttta gctataatta
     6001 tagacgatgc ccttgtatcc cattctgtct cgcttgcccc tactttttat cttttatata
     6061 ccataatgaa cgctgccgct actaaccata ccccgatttt acatttcgga ctcccaagga
     6121 cgtacaaaat agaaaactat agaaaaaaat aatcagaaaa tagcatgtca tctctttgta
     6181 aaacgcgttt gcaagaagaa aggaaacaat ggagaagaga tcatccattt gtatgtaaat
     6241 tttagtaaac ttgaagaaat cactaacaac ttctcttact tagggattct atgcaaaacc
     6301 ttgtaaatca tctgatggag gactcgattt aatgaattgg aaggttggat ttccaggtgg
     6361 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6421 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6481 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6541 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6601 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6661 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6721 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6781 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6841 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6901 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6961 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7021 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7081 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7141 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7201 atctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7261 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7321 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7381 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7441 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7501 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7561 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7621 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     7681 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7741 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7801 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7861 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc
     7921 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7981 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8041 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8101 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     8161 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     8221 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     8281 ggaag
//
