LOCUS       Exported                8278 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 8278)
  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..8278
                     /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..4116
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     terminator      4143..5134
                     /label=nmt1 terminator
     rep_origin      5214..5997
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6373..6477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6478..7338
                     /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      7509..8097
                     /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 ggaagaggaa tcctggcata tcatcaattg aataagttga attaattatt tcaatctcat
     4081 tctcactttc tgacttatag tcgctttgtt aaatcatatg tcgactctag aggatccccg
     4141 ggtaaaagga atgtctccct tgccagtact gctagggttt ttctttcaaa ctatggaagc
     4201 ccattcaagc tgcatattac gattttgttt ttcgctttta gaaagtggtt tagatgagat
     4261 aatagaaaaa ttcttgatct ccgacaacga gtacttttat tttttttgct aatcacttta
     4321 ctcaatatta gctcgaaatc gtagaaacgt agacgggtgc gggataccga gtggtgtagt
     4381 taagaatttt tataaaccac gtggcccaaa aatatgaacc caaaacgttt atacatgagt
     4441 atactttaag aaggctatac cccttcgtgt tagatgtagt tttagctacc caacccgagt
     4501 ctatgagctt gacttcagat gtagaaggca ttaaatcgtt ttgaatatta attaaaaaac
     4561 gatgaaaatt aaatatttaa aagcaatcat acgctgaaaa tttagtgctg tggctaatcc
     4621 ttcaacatgg aaatgccata aaagtgactt tgacaaaaaa aaaagtatat acaggtagta
     4681 aactcatcta cttcattgac tttgtttaca gcatgtggaa ggaggaatat ttattgctaa
     4741 atcgtagttt aacattcaat aagtaatact attgaaattc gacaagattg gccgcatgga
     4801 tgaaaaagag gcattttgct ttgggagaat tagttcaaat tagaactgaa aaaaaaaact
     4861 ttacgaggca aaaatgtcgg attgagatcg taaaagttcg ctcgtcgtct tttgctttgt
     4921 gattgttttc atggatacat cttgctggat atttaaattt tagtactatg tataagatat
     4981 tctataaatg ttttatcacc caaacctgtt agcgccttct taattctatt caatctggct
     5041 tttgctctga gactacttct tggactttca ctacttgtta gttatacgga atttgtgtaa
     5101 ttagaagtga aataatcctt tctattagta atgcgagctg caattcgagt ctaactcctt
     5161 aaccactcgg acatagtgac ttatctgaca ctcattgtaa attaatatat atataggcat
     5221 tttgtttagt taaaggtact taagtaatta gtataaacga accaatttta taatcaggaa
     5281 gttaagtgaa tggtagcaca tgtcgtaaaa attgtgaatt tttattgaat aatattttaa
     5341 atacaagcct ttctagacta ggtatactca taaacatata tgagcaaaag gatagaggag
     5401 attacattgc atcttctaca aatttattta ttgcccttta ctgaaaaatt aaataatgag
     5461 tactaaatga taaaaagcgc tcagtacaag aaagattgca aaaatattgc attcttcatg
     5521 aattaaagtt gcatataagg catattgaaa gtaatagtac taaaacagca gttagcgaaa
     5581 attaatagaa ttatattcgc aagacaattg tgacattaaa ttaaaaaatt gtagaatttt
     5641 tactatcctc tttaacgcca tgagccttta taaaaaggtt aaattagttt taacattctt
     5701 tttttgagta agagtttaca atttatcaaa acctgtgtta ttatattcat tagtttcaat
     5761 ttattagcat ctagagaaaa atcaattggc agttacattg ttggaattta tgaagaaaaa
     5821 gaatctacaa cggagaataa gttgctgatc gctttcccta aaattgtata ttttgctgag
     5881 cttattttga catttcgttg aagttttctc taattcgcat tcattttaag taaaacaatg
     5941 agaaataaaa ttacaaaaaa tacaaattaa aatacaattt ttagctataa ttatagacga
     6001 tgcccttgta tcccattctg tctcgcttgc ccctactttt tatcttttat ataccataat
     6061 gaacgctgcc gctactaacc ataccccgat tttacatttc ggactcccaa ggacgtacaa
     6121 aatagaaaac tatagaaaaa aataatcaga aaatagcatg tcatctcttt gtaaaacgcg
     6181 tttgcaagaa gaaaggaaac aatggagaag agatcatcca tttgtatgta aattttagta
     6241 aacttgaaga aatcactaac aacttctctt acttagggat tctatgcaaa accttgtaaa
     6301 tcatctgatg gaggactcga tttaatgaat tggaaggttg gatttccagg tggcactttt
     6361 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6421 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6481 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6541 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6601 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6661 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6721 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6781 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6841 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6901 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6961 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     7021 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7081 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7141 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggatctcgc
     7201 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7261 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7321 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7381 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7441 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7501 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7561 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7621 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     7681 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7741 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7801 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7861 cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt
     7921 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7981 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     8041 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     8101 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     8161 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     8221 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
