LOCUS       Exported                6417 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 6417)
  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..6417
                     /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    1035..1040
                     /label=BsaI
                     /label=BsmB1
     misc_feature    1061..1066
                     /label=BsaI
     promoter        1098..2255
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     terminator      2282..3273
                     /label=nmt1 terminator
     rep_origin      3353..4136
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        4512..4616
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4617..5477
                     /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      5648..6236
                     /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 tttagagacc ggttctttgg gtcaagaccg ggtctctgtc gatcgactct
     1081 agaggatcag aaaattatcg ccataaaaga cagaataagt catcagcggt tgtttcattt
     1141 cctatatttt ttttttattt ttttattttt taataaggga aaatttaacg tctaaggata
     1201 cagaagattg ttagcacatt aaagtaataa aggcttaagt agtaagtgcc ttagcatgtt
     1261 attgtatttc aaaggacata atctaaaata ataacaatat catttctcac aagttattca
     1321 attttctttt ttttttctaa taatatcaag aatgtattat ttgtttgaca taagtcaact
     1381 aatttattta atatgctgga ttaatcttgc agacatgtaa attaacaagt tttagtcaaa
     1441 taacgttgaa gtttcaatga actcaaataa tttctctttt tttttatata accataagtc
     1501 taatctgatt tatattttcc gcagggatca actgaagtta tgacatttgg attggatcac
     1561 ttataacctt ggtcgccaaa taatacaaaa atcagcgtta taaaacaaag aaggtttttg
     1621 ttaagaaatt aatcctcttt cttgataaga aagttgaacc gaaattgcag atactgatat
     1681 atgaaaataa tacccacaat tttgggaata gcgcaagcct caatttaaac aataggtgag
     1741 gacacatgat aatgacctca atgattgtta gaagaaaaga gcctcattac aaaatcgaaa
     1801 aatgaatggt tgggtacaag tttccaaaac atggtaaagt ggactttgcg tatgagacgt
     1861 aaatagaaaa aaacacttgt tatatgtttt ctagaattat tgttgtctct ttatggttgg
     1921 atgatgcaaa atagtaattt cggttagttg ctgtaaaaca ccacgagaca aatagatatg
     1981 gatatttatt aaatcaggaa aaacgtaact ctcggctact ggatggttca gtcacccaac
     2041 gattactggg gagagaaaac agggcaaaag caaagcttaa aggaatccga ttgtcattcg
     2101 gcaatgtgca gcgaaactaa aaaccggata atggacctgt taatcgaaac attgaagatg
     2161 gaagaggaat cctggcatat catcaattga ataagttgaa ttaattattt caatctcatt
     2221 ctcactttct gacttatagt cgctttgtta aatcatatgt cgactctaga ggatccccgg
     2281 gtaaaaggaa tgtctccctt gccagtactg ctagggtttt tctttcaaac tatggaagcc
     2341 cattcaagct gcatattacg attttgtttt tcgcttttag aaagtggttt agatgagata
     2401 atagaaaaat tcttgatctc cgacaacgag tacttttatt ttttttgcta atcactttac
     2461 tcaatattag ctcgaaatcg tagaaacgta gacgggtgcg ggataccgag tggtgtagtt
     2521 aagaattttt ataaaccacg tggcccaaaa atatgaaccc aaaacgttta tacatgagta
     2581 tactttaaga aggctatacc ccttcgtgtt agatgtagtt ttagctaccc aacccgagtc
     2641 tatgagcttg acttcagatg tagaaggcat taaatcgttt tgaatattaa ttaaaaaacg
     2701 atgaaaatta aatatttaaa agcaatcata cgctgaaaat ttagtgctgt ggctaatcct
     2761 tcaacatgga aatgccataa aagtgacttt gacaaaaaaa aaagtatata caggtagtaa
     2821 actcatctac ttcattgact ttgtttacag catgtggaag gaggaatatt tattgctaaa
     2881 tcgtagttta acattcaata agtaatacta ttgaaattcg acaagattgg ccgcatggat
     2941 gaaaaagagg cattttgctt tgggagaatt agttcaaatt agaactgaaa aaaaaaactt
     3001 tacgaggcaa aaatgtcgga ttgagatcgt aaaagttcgc tcgtcgtctt ttgctttgtg
     3061 attgttttca tggatacatc ttgctggata tttaaatttt agtactatgt ataagatatt
     3121 ctataaatgt tttatcaccc aaacctgtta gcgccttctt aattctattc aatctggctt
     3181 ttgctctgag actacttctt ggactttcac tacttgttag ttatacggaa tttgtgtaat
     3241 tagaagtgaa ataatccttt ctattagtaa tgcgagctgc aattcgagtc taactcctta
     3301 accactcgga catagtgact tatctgacac tcattgtaaa ttaatatata tataggcatt
     3361 ttgtttagtt aaaggtactt aagtaattag tataaacgaa ccaattttat aatcaggaag
     3421 ttaagtgaat ggtagcacat gtcgtaaaaa ttgtgaattt ttattgaata atattttaaa
     3481 tacaagcctt tctagactag gtatactcat aaacatatat gagcaaaagg atagaggaga
     3541 ttacattgca tcttctacaa atttatttat tgccctttac tgaaaaatta aataatgagt
     3601 actaaatgat aaaaagcgct cagtacaaga aagattgcaa aaatattgca ttcttcatga
     3661 attaaagttg catataaggc atattgaaag taatagtact aaaacagcag ttagcgaaaa
     3721 ttaatagaat tatattcgca agacaattgt gacattaaat taaaaaattg tagaattttt
     3781 actatcctct ttaacgccat gagcctttat aaaaaggtta aattagtttt aacattcttt
     3841 ttttgagtaa gagtttacaa tttatcaaaa cctgtgttat tatattcatt agtttcaatt
     3901 tattagcatc tagagaaaaa tcaattggca gttacattgt tggaatttat gaagaaaaag
     3961 aatctacaac ggagaataag ttgctgatcg ctttccctaa aattgtatat tttgctgagc
     4021 ttattttgac atttcgttga agttttctct aattcgcatt cattttaagt aaaacaatga
     4081 gaaataaaat tacaaaaaat acaaattaaa atacaatttt tagctataat tatagacgat
     4141 gcccttgtat cccattctgt ctcgcttgcc cctacttttt atcttttata taccataatg
     4201 aacgctgccg ctactaacca taccccgatt ttacatttcg gactcccaag gacgtacaaa
     4261 atagaaaact atagaaaaaa ataatcagaa aatagcatgt catctctttg taaaacgcgt
     4321 ttgcaagaag aaaggaaaca atggagaaga gatcatccat ttgtatgtaa attttagtaa
     4381 acttgaagaa atcactaaca acttctctta cttagggatt ctatgcaaaa ccttgtaaat
     4441 catctgatgg aggactcgat ttaatgaatt ggaaggttgg atttccaggt ggcacttttc
     4501 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4561 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4621 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4681 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4741 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4801 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4861 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4921 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4981 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5041 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5101 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5161 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5221 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5281 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggatctcgcg
     5341 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5401 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5461 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5521 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5581 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5641 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5701 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5761 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     5821 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5881 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5941 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6001 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc
     6061 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6121 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6181 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6241 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6301 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6361 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
