LOCUS       Exported                8573 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 8573)
  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..8573
                     /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(1034..1183)
                     /label=150bp lys9 downstream
     misc_feature    complement(1184..2831)
                     /label=lys9
     CDS             complement(1317..2669)
                     /codon_start=1
                     /translation="MPSILLLGSGFVAHPTLEYLSRRKENNITVACRTLSKAEAFINGI
                     PNSKAIALDVNDEAALEKAVSEHDLTISLIPYTYHATVMKAAIKHGKHVCTTSYVNPKM
                     AELEEAAIKAGSICMNEIGVDPGIDHLYAIKTIEEVHKAGGKIKSFLSYCGGLPAPEDS
                     NNPLGYKFSWSSRGVLLALRNSAKFYENGKLVEIDGKDLMETAKPYFIYPGYAFVCYPN
                     RDSTVYQERYQIPEAETIIRGTLRYQGFPEFIHCLVDMGFLDETAQEYLSPEAPALPWK
                     EVTARVIKAESSSEADLIKKISSIHKFKDEDDKKRILNGLKWLGMFSSKPVTPRGNPLD
                     TLCATLEELMQYEEGERDMLILQHKFEVETKEGKRQTRTCTLLDYGVPNGYTSMAKLVG
                     VPCGVATQQILDGVINTPGVLAPNDMKLCGPLIDTLAKEGIRLEEEIIDEE"
     misc_feature    complement(2832..3216)
                     /label=400bp lys9 upstream
     promoter        3247..4411
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     terminator      4438..5429
                     /label=nmt1 terminator
     rep_origin      5509..6292
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6668..6772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6773..7633
                     /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      7804..8392
                     /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 ttttgttcaa ataatcgaga agaggttatt ttttcttcga gttttacgtc
     1081 ggttacctaa gactaaatac atctacttag aaaatacgaa aaccccaagt aaagcttcac
     1141 ataaataagg ttgcaaaaca aactgaacat ttatacttat gaacatgtag ataaattgcc
     1201 aaaaaaaagg ttgacgaact ctaagattac gaataattat agaaaatgaa gaagctttgg
     1261 tgttctaaaa tgtcaaactt atatataaga gtttggaacc aaagcgtgaa acatgcttac
     1321 tcttcgtcaa taatttcttc ctccaatcga ataccttctt tggcaagtgt atcaatcaat
     1381 ggaccgcaaa gcttcatatc attaggagcc aaaacacctg gggtgttaat aacaccatca
     1441 agaatttgct gagtagcaac accacaagga acgccgacta attttgccat agaggtgtag
     1501 ccattaggaa caccgtagtc aagcaaagtg caagtacgag tttggcgttt tccttctttg
     1561 gtttcaactt caaacttgtg ttgaaggatt aacatatcac gttcaccttc ttcgtattgc
     1621 ataagttcct ccaaggtagc gcataaggtg tccaaaggat taccacgagg agtaacgggt
     1681 ttgctagaga acataccaag ccacttcaaa ccattcaaaa tgcgcttctt gtcgtcttca
     1741 tctttgaatt tgtggatgga agaaattttc ttgattaggt cggcttcgga ggaagattca
     1801 gccttgatga cacgagcagt aacctccttc caaggcaaag cgggtgcttc aggagataag
     1861 tattcctgag ctgtctcatc caaaaatccc atatcaacca aacaatgaat aaactcggga
     1921 aatccttggt aacgtaaagt accgcgaatg atagtttctg cttcaggtat ttgatagcgc
     1981 tcttggtaga cagtggaatc acgattagga taacaaacga aagcgtaacc gggatagatg
     2041 aaatagggtt tagcagtttc catcaaatcc ttgccgtcaa tttcaacaag tttgccgttt
     2101 tcataaaact tggcagagtt gcgaagagcc aacaaaacac cacgagaaga ccaagaaaac
     2161 ttgtatccta aaggattgtt ggagtcctcg ggagcgggaa gacctccaca ataagacaag
     2221 aaagacttaa tcttaccgcc agctttatga acctcttcaa tggtcttgat agcatataaa
     2281 tgatcaatac cgggatcgac gccgatttcg ttcatacaaa tagaaccagc cttgatggca
     2341 gcttcctcca actcagccat tttgggattc acataactgg tggtacaaac gtgtttacca
     2401 tgcttaatgg cagctttcat aactgttgca tgataagtgt aaggaatcaa gctgatggta
     2461 agatcatgtt cggaaacagc tttctcaaga gctgcttcat cattgacatc caaagcaatg
     2521 gctttgctgt ttgggatacc attaataaat gcttcagcct tagaaagagt tctgcatgct
     2581 acggttatat tattttcttt acgacgagat aaatattcta aagtaggatg agcgacaaat
     2641 cccgaaccaa gtaataagat ggaaggcatt ttgatgttga aaatgaaaga ggcagtcagt
     2701 taaggacaag tggtaaacca aagacttctg ttttgtcgtc acggttttag cgatatataa
     2761 ggggttcctt tttatccaca gatttatttc tgatggccga aatcgtcgtc taaattgatg
     2821 aaaactgatt atctcgaatc ggttaaaaaa gcagtcacat ggaccatttt ttgaattaat
     2881 atttaatcgt tacgtttcct ctcaatgtta tgtttggctt actaggggaa ttggttatta
     2941 tcgacaagtt tagtcgataa tttgttagca gcgatgtcgc aactgatagt tttagcaact
     3001 ttttttacgt agtttttttc tgcacaacag aagttgctta aaagctagta gatgccattc
     3061 gttgttgttt gggggtagat taaagaattg taagctttag tataagaata atttgtttct
     3121 cctctttctg cctatctgtt gttcataatc aactgcgtag ttctggtgaa ggagtaaatt
     3181 aaactgcctc gcaccgccgt actcattttt acctcggtcg atcgactcta gaggatcaga
     3241 aaattatcgc cataaaagac agaataagtc atcagcggtt gtttcatttc ctatattttt
     3301 tttttatttt tttatttttt aataagggaa aatttaacgt ctaaggatac agaagattgt
     3361 tagcacatta aagtaataaa ggcttaagta gtaagtgcct tagcatgtta ttgtatttca
     3421 aaggacataa tctaaaataa taacaatatc atttctcaca agttattcaa ttttcttttt
     3481 tttttctaat aatatcaaga atgtattatt tgtttgacat aagtcaacta atttatttaa
     3541 tatgctggat taatcttgca gacatgtaaa ttaacaagtt ttagtcaaat aacgttgaag
     3601 tttcaatgaa ctcaaataat ttctcttttt ttttatataa ccataagtct aatctgattt
     3661 atattttccg cagggatcaa ctgaagttat gacatttgga ttggatcact tataaccttg
     3721 gtcgccaaat aatacaaaaa tcagcgttat aaaacaaaga aggtttttgt taagaaatta
     3781 atcctctttc ttgataagaa agttgaaccg aaattgcaga tactgatata tgaaaataat
     3841 acccacaatt ttgggaatag cgcaagcctc aatttaaaca ataggtgagg acacatgata
     3901 atgacctcaa tgattgttag aagaaaagag cctcattaca aaatcgaaaa atgaatggtt
     3961 gggtacaagt ttccaaaaca tggtaaagtg gactttgcgt atgagacgta aatagaaaaa
     4021 aacacttgtt atatgttttc tagaattatt gttgtctctt tatggttgga tgatgcaaaa
     4081 tagtaatttc ggttagttgc tgtaaaacac cacgagacaa atagatatgg atatttatta
     4141 aatcaggaaa aacgtaactc tcggctactg gatggttcag tcacccaacg attactgggg
     4201 agagaaaaca gggcaaaagc aaagcttaaa ggaatccgat tgtcattcgg caatgtgcag
     4261 cgaaactaaa aaccggataa tggacctgtt aatcgaaaca ttgaagatat ataaaggaag
     4321 aggaatcctg gcatatcatc aattgaataa gttgaattaa ttatttcaat ctcattctca
     4381 ctttctgact tatagtcgct ttgttaaatc atatgtcgac tctagaggat ccccgggtaa
     4441 aaggaatgtc tcccttgcca gtactgctag ggtttttctt tcaaactatg gaagcccatt
     4501 caagctgcat attacgattt tgtttttcgc ttttagaaag tggtttagat gagataatag
     4561 aaaaattctt gatctccgac aacgagtact tttatttttt ttgctaatca ctttactcaa
     4621 tattagctcg aaatcgtaga aacgtagacg ggtgcgggat accgagtggt gtagttaaga
     4681 atttttataa accacgtggc ccaaaaatat gaacccaaaa cgtttataca tgagtatact
     4741 ttaagaaggc tatacccctt cgtgttagat gtagttttag ctacccaacc cgagtctatg
     4801 agcttgactt cagatgtaga aggcattaaa tcgttttgaa tattaattaa aaaacgatga
     4861 aaattaaata tttaaaagca atcatacgct gaaaatttag tgctgtggct aatccttcaa
     4921 catggaaatg ccataaaagt gactttgaca aaaaaaaaag tatatacagg tagtaaactc
     4981 atctacttca ttgactttgt ttacagcatg tggaaggagg aatatttatt gctaaatcgt
     5041 agtttaacat tcaataagta atactattga aattcgacaa gattggccgc atggatgaaa
     5101 aagaggcatt ttgctttggg agaattagtt caaattagaa ctgaaaaaaa aaactttacg
     5161 aggcaaaaat gtcggattga gatcgtaaaa gttcgctcgt cgtcttttgc tttgtgattg
     5221 ttttcatgga tacatcttgc tggatattta aattttagta ctatgtataa gatattctat
     5281 aaatgtttta tcacccaaac ctgttagcgc cttcttaatt ctattcaatc tggcttttgc
     5341 tctgagacta cttcttggac tttcactact tgttagttat acggaatttg tgtaattaga
     5401 agtgaaataa tcctttctat tagtaatgcg agctgcaatt cgagtctaac tccttaacca
     5461 ctcggacata gtgacttatc tgacactcat tgtaaattaa tatatatata ggcattttgt
     5521 ttagttaaag gtacttaagt aattagtata aacgaaccaa ttttataatc aggaagttaa
     5581 gtgaatggta gcacatgtcg taaaaattgt gaatttttat tgaataatat tttaaataca
     5641 agcctttcta gactaggtat actcataaac atatatgagc aaaaggatag aggagattac
     5701 attgcatctt ctacaaattt atttattgcc ctttactgaa aaattaaata atgagtacta
     5761 aatgataaaa agcgctcagt acaagaaaga ttgcaaaaat attgcattct tcatgaatta
     5821 aagttgcata taaggcatat tgaaagtaat agtactaaaa cagcagttag cgaaaattaa
     5881 tagaattata ttcgcaagac aattgtgaca ttaaattaaa aaattgtaga atttttacta
     5941 tcctctttaa cgccatgagc ctttataaaa aggttaaatt agttttaaca ttcttttttt
     6001 gagtaagagt ttacaattta tcaaaacctg tgttattata ttcattagtt tcaatttatt
     6061 agcatctaga gaaaaatcaa ttggcagtta cattgttgga atttatgaag aaaaagaatc
     6121 tacaacggag aataagttgc tgatcgcttt ccctaaaatt gtatattttg ctgagcttat
     6181 tttgacattt cgttgaagtt ttctctaatt cgcattcatt ttaagtaaaa caatgagaaa
     6241 taaaattaca aaaaatacaa attaaaatac aatttttagc tataattata gacgatgccc
     6301 ttgtatccca ttctgtctcg cttgccccta ctttttatct tttatatacc ataatgaacg
     6361 ctgccgctac taaccatacc ccgattttac atttcggact cccaaggacg tacaaaatag
     6421 aaaactatag aaaaaaataa tcagaaaata gcatgtcatc tctttgtaaa acgcgtttgc
     6481 aagaagaaag gaaacaatgg agaagagatc atccatttgt atgtaaattt tagtaaactt
     6541 gaagaaatca ctaacaactt ctcttactta gggattctat gcaaaacctt gtaaatcatc
     6601 tgatggagga ctcgatttaa tgaattggaa ggttggattt ccaggtggca cttttcgggg
     6661 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6721 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6781 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6841 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6901 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6961 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7021 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7081 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7141 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7201 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7261 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7321 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7381 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7441 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggat ctcgcggtat
     7501 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7561 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7621 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7681 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7741 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7801 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7861 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7921 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     7981 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     8041 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     8101 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     8161 gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga
     8221 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     8281 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     8341 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     8401 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     8461 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     8521 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
