LOCUS       Exported                8281 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 8281)
  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..8281
                     /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..4119
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     terminator      4146..5137
                     /label=nmt1 terminator
     rep_origin      5217..6000
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6376..6480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6481..7341
                     /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      7512..8100
                     /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 aaaggaagag gaatcctggc atatcatcaa ttgaataagt tgaattaatt atttcaatct
     4081 cattctcact ttctgactta tagtcgcttt gttaaatcat atgtcgactc tagaggatcc
     4141 ccgggtaaaa ggaatgtctc ccttgccagt actgctaggg tttttctttc aaactatgga
     4201 agcccattca agctgcatat tacgattttg tttttcgctt ttagaaagtg gtttagatga
     4261 gataatagaa aaattcttga tctccgacaa cgagtacttt tatttttttt gctaatcact
     4321 ttactcaata ttagctcgaa atcgtagaaa cgtagacggg tgcgggatac cgagtggtgt
     4381 agttaagaat ttttataaac cacgtggccc aaaaatatga acccaaaacg tttatacatg
     4441 agtatacttt aagaaggcta taccccttcg tgttagatgt agttttagct acccaacccg
     4501 agtctatgag cttgacttca gatgtagaag gcattaaatc gttttgaata ttaattaaaa
     4561 aacgatgaaa attaaatatt taaaagcaat catacgctga aaatttagtg ctgtggctaa
     4621 tccttcaaca tggaaatgcc ataaaagtga ctttgacaaa aaaaaaagta tatacaggta
     4681 gtaaactcat ctacttcatt gactttgttt acagcatgtg gaaggaggaa tatttattgc
     4741 taaatcgtag tttaacattc aataagtaat actattgaaa ttcgacaaga ttggccgcat
     4801 ggatgaaaaa gaggcatttt gctttgggag aattagttca aattagaact gaaaaaaaaa
     4861 actttacgag gcaaaaatgt cggattgaga tcgtaaaagt tcgctcgtcg tcttttgctt
     4921 tgtgattgtt ttcatggata catcttgctg gatatttaaa ttttagtact atgtataaga
     4981 tattctataa atgttttatc acccaaacct gttagcgcct tcttaattct attcaatctg
     5041 gcttttgctc tgagactact tcttggactt tcactacttg ttagttatac ggaatttgtg
     5101 taattagaag tgaaataatc ctttctatta gtaatgcgag ctgcaattcg agtctaactc
     5161 cttaaccact cggacatagt gacttatctg acactcattg taaattaata tatatatagg
     5221 cattttgttt agttaaaggt acttaagtaa ttagtataaa cgaaccaatt ttataatcag
     5281 gaagttaagt gaatggtagc acatgtcgta aaaattgtga atttttattg aataatattt
     5341 taaatacaag cctttctaga ctaggtatac tcataaacat atatgagcaa aaggatagag
     5401 gagattacat tgcatcttct acaaatttat ttattgccct ttactgaaaa attaaataat
     5461 gagtactaaa tgataaaaag cgctcagtac aagaaagatt gcaaaaatat tgcattcttc
     5521 atgaattaaa gttgcatata aggcatattg aaagtaatag tactaaaaca gcagttagcg
     5581 aaaattaata gaattatatt cgcaagacaa ttgtgacatt aaattaaaaa attgtagaat
     5641 ttttactatc ctctttaacg ccatgagcct ttataaaaag gttaaattag ttttaacatt
     5701 ctttttttga gtaagagttt acaatttatc aaaacctgtg ttattatatt cattagtttc
     5761 aatttattag catctagaga aaaatcaatt ggcagttaca ttgttggaat ttatgaagaa
     5821 aaagaatcta caacggagaa taagttgctg atcgctttcc ctaaaattgt atattttgct
     5881 gagcttattt tgacatttcg ttgaagtttt ctctaattcg cattcatttt aagtaaaaca
     5941 atgagaaata aaattacaaa aaatacaaat taaaatacaa tttttagcta taattataga
     6001 cgatgccctt gtatcccatt ctgtctcgct tgcccctact ttttatcttt tatataccat
     6061 aatgaacgct gccgctacta accatacccc gattttacat ttcggactcc caaggacgta
     6121 caaaatagaa aactatagaa aaaaataatc agaaaatagc atgtcatctc tttgtaaaac
     6181 gcgtttgcaa gaagaaagga aacaatggag aagagatcat ccatttgtat gtaaatttta
     6241 gtaaacttga agaaatcact aacaacttct cttacttagg gattctatgc aaaaccttgt
     6301 aaatcatctg atggaggact cgatttaatg aattggaagg ttggatttcc aggtggcact
     6361 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6421 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6481 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6541 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6601 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6661 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6721 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6781 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6841 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6901 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6961 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7021 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7081 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7141 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggatct
     7201 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7261 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7321 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7381 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7441 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7501 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7561 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7621 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     7681 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7741 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7801 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7861 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
     7921 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7981 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8041 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8101 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8161 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     8221 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     8281 g
//
