LOCUS       Exported                8141 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 8141)
  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..8141
                     /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"
     CDS             complement(1469..2263)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     promoter        2822..3979
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     terminator      4006..4997
                     /label=nmt1 terminator
     rep_origin      5077..5860
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6236..6340
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6341..7201
                     /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      7372..7960
                     /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 tttagcttgt gatattgacg aaactttttg acatctaatt tattctgttc
     1081 caacaccaat gtttataacc aagttttatc ttgtttgtct acatggtatt ttacattcat
     1141 ctacatacat ctttcattgg ctttgtacat agttatcatt acaagtctaa aaaaattcac
     1201 tcttttctta ttcaatgtca atccaagaga aaagattgtg gtaatgttgt aggagcatgt
     1261 ttaataaatt actatagcaa attacttttt attcccaagg tgtttatcta taatagttaa
     1321 tattttagtc gctacataaa attttaccaa agagtacttg tatactaatt ctaaatgcct
     1381 tctgacataa aacgcctagg aaaacaaacg caaacaaggc atcgactttt tcaataacca
     1441 accaaaaaaa ttttacatta gtcttttttt aatgctgaga aagtctttgc tgatatgcct
     1501 tccaaccagc ttctctatat ctcttggctt cgacaacagg attacgacca gctccataga
     1561 ctccacgacc aacaatgatg atatcgctac cgcagtttac aatcacttct tcaggagtac
     1621 gatattgctg tcccagcccg tctcctttaa catccaagcc gataccaggg gacatagtta
     1681 tgtagtcgct ttgaaggtta ggaaatcgac gaccagctat aaagccaaag caaaaatcgg
     1741 tatgcttctc aaaccattct aaggttttct ctgtgtagga accagtagcc aaagagcctt
     1801 tggaagacat ttcagccaaa agcaagagtc cacgtcccaa aggtaaacca acttctttga
     1861 ggccttgtat aataccctcg cctggcactg tatggcaatt tgtgatatga gcccaagaag
     1921 caattttgta cacaccagat gcatattgta gcttgacggt atttccaatg tctgcgaatt
     1981 tgcgatcctc aaagataaga aaacgatgct ttttacctaa ggccaccagt ttttctacca
     2041 tatcctggtc gaaatcctcg acaacgtcaa tatgtgtctt gataacacag acatagggtc
     2101 caattttatc taccaattct aagatttcgg atttcttcgt caaatcgacc gcgactgaca
     2161 agttgctttg cttttcttcc atcaaagcca acaattcctt ggcaatggga tttttcatcc
     2221 cctcagctct agctgaatag ctttgaaata ctctagcatc cataactttg cttttaaacc
     2281 tttaatttcg atccaagcaa aaaagaggtt cttggtagga caatacggta agaaaacacg
     2341 acatgtgcag agatgccgac gaagcatagt taaactggga tggtaaaatc aattaagaat
     2401 ttataaagac aaaattgtat aagtctctaa aacatcttaa ttatacctca cagaactatc
     2461 taaaatatat tcacaaagtg caaacattat catgaaaaag aaccatttta atttaaagca
     2521 agggcattaa ggcttattta cagaatttct tacttttgta aagattataa ggctgattat
     2581 ctttttcacc atgccaaaaa ttacacaaga tagaatggat gtttgaaatt aaacgtgagt
     2641 atacaaacaa atacactagg taaatcgaaa catttttttc tccattaagt aacaaattcc
     2701 tatttagaga aagaatgctg agtagattaa ataatctata caaacttttt taacacaaat
     2761 gcatacatat agccagtggg atttgtagct agtcgatcga ctctagagga tcagaaaatt
     2821 atcgccataa aagacagaat aagtcatcag cggttgtttc atttcctata tttttttttt
     2881 atttttttat tttttaataa gggaaaattt aacgtctaag gatacagaag attgttagca
     2941 cattaaagta ataaaggctt aagtagtaag tgccttagca tgttattgta tttcaaagga
     3001 cataatctaa aataataaca atatcatttc tcacaagtta ttcaattttc tttttttttt
     3061 ctaataatat caagaatgta ttatttgttt gacataagtc aactaattta tttaatatgc
     3121 tggattaatc ttgcagacat gtaaattaac aagttttagt caaataacgt tgaagtttca
     3181 atgaactcaa ataatttctc ttttttttta tataaccata agtctaatct gatttatatt
     3241 ttccgcaggg atcaactgaa gttatgacat ttggattgga tcacttataa ccttggtcgc
     3301 caaataatac aaaaatcagc gttataaaac aaagaaggtt tttgttaaga aattaatcct
     3361 ctttcttgat aagaaagttg aaccgaaatt gcagatactg atatatgaaa ataataccca
     3421 caattttggg aatagcgcaa gcctcaattt aaacaatagg tgaggacaca tgataatgac
     3481 ctcaatgatt gttagaagaa aagagcctca ttacaaaatc gaaaaatgaa tggttgggta
     3541 caagtttcca aaacatggta aagtggactt tgcgtatgag acgtaaatag aaaaaaacac
     3601 ttgttatatg ttttctagaa ttattgttgt ctctttatgg ttggatgatg caaaatagta
     3661 atttcggtta gttgctgtaa aacaccacga gacaaataga tatggatatt tattaaatca
     3721 ggaaaaacgt aactctcggc tactggatgg ttcagtcacc caacgattac tggggagaga
     3781 aaacagggca aaagcaaagc ttaaaggaat ccgattgtca ttcggcaatg tgcagcgaaa
     3841 ctaaaaaccg gataatggac ctgttaatcg aaacattgaa gatggaagag gaatcctggc
     3901 atatcatcaa ttgaataagt tgaattaatt atttcaatct cattctcact ttctgactta
     3961 tagtcgcttt gttaaatcat atgtcgactc tagaggatcc ccgggtaaaa ggaatgtctc
     4021 ccttgccagt actgctaggg tttttctttc aaactatgga agcccattca agctgcatat
     4081 tacgattttg tttttcgctt ttagaaagtg gtttagatga gataatagaa aaattcttga
     4141 tctccgacaa cgagtacttt tatttttttt gctaatcact ttactcaata ttagctcgaa
     4201 atcgtagaaa cgtagacggg tgcgggatac cgagtggtgt agttaagaat ttttataaac
     4261 cacgtggccc aaaaatatga acccaaaacg tttatacatg agtatacttt aagaaggcta
     4321 taccccttcg tgttagatgt agttttagct acccaacccg agtctatgag cttgacttca
     4381 gatgtagaag gcattaaatc gttttgaata ttaattaaaa aacgatgaaa attaaatatt
     4441 taaaagcaat catacgctga aaatttagtg ctgtggctaa tccttcaaca tggaaatgcc
     4501 ataaaagtga ctttgacaaa aaaaaaagta tatacaggta gtaaactcat ctacttcatt
     4561 gactttgttt acagcatgtg gaaggaggaa tatttattgc taaatcgtag tttaacattc
     4621 aataagtaat actattgaaa ttcgacaaga ttggccgcat ggatgaaaaa gaggcatttt
     4681 gctttgggag aattagttca aattagaact gaaaaaaaaa actttacgag gcaaaaatgt
     4741 cggattgaga tcgtaaaagt tcgctcgtcg tcttttgctt tgtgattgtt ttcatggata
     4801 catcttgctg gatatttaaa ttttagtact atgtataaga tattctataa atgttttatc
     4861 acccaaacct gttagcgcct tcttaattct attcaatctg gcttttgctc tgagactact
     4921 tcttggactt tcactacttg ttagttatac ggaatttgtg taattagaag tgaaataatc
     4981 ctttctatta gtaatgcgag ctgcaattcg agtctaactc cttaaccact cggacatagt
     5041 gacttatctg acactcattg taaattaata tatatatagg cattttgttt agttaaaggt
     5101 acttaagtaa ttagtataaa cgaaccaatt ttataatcag gaagttaagt gaatggtagc
     5161 acatgtcgta aaaattgtga atttttattg aataatattt taaatacaag cctttctaga
     5221 ctaggtatac tcataaacat atatgagcaa aaggatagag gagattacat tgcatcttct
     5281 acaaatttat ttattgccct ttactgaaaa attaaataat gagtactaaa tgataaaaag
     5341 cgctcagtac aagaaagatt gcaaaaatat tgcattcttc atgaattaaa gttgcatata
     5401 aggcatattg aaagtaatag tactaaaaca gcagttagcg aaaattaata gaattatatt
     5461 cgcaagacaa ttgtgacatt aaattaaaaa attgtagaat ttttactatc ctctttaacg
     5521 ccatgagcct ttataaaaag gttaaattag ttttaacatt ctttttttga gtaagagttt
     5581 acaatttatc aaaacctgtg ttattatatt cattagtttc aatttattag catctagaga
     5641 aaaatcaatt ggcagttaca ttgttggaat ttatgaagaa aaagaatcta caacggagaa
     5701 taagttgctg atcgctttcc ctaaaattgt atattttgct gagcttattt tgacatttcg
     5761 ttgaagtttt ctctaattcg cattcatttt aagtaaaaca atgagaaata aaattacaaa
     5821 aaatacaaat taaaatacaa tttttagcta taattataga cgatgccctt gtatcccatt
     5881 ctgtctcgct tgcccctact ttttatcttt tatataccat aatgaacgct gccgctacta
     5941 accatacccc gattttacat ttcggactcc caaggacgta caaaatagaa aactatagaa
     6001 aaaaataatc agaaaatagc atgtcatctc tttgtaaaac gcgtttgcaa gaagaaagga
     6061 aacaatggag aagagatcat ccatttgtat gtaaatttta gtaaacttga agaaatcact
     6121 aacaacttct cttacttagg gattctatgc aaaaccttgt aaatcatctg atggaggact
     6181 cgatttaatg aattggaagg ttggatttcc aggtggcact tttcggggaa atgtgcgcgg
     6241 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6301 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6361 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6421 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6481 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6541 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6601 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6661 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6721 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6781 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6841 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6901 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6961 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7021 gtttattgct gataaatctg gagccggtga gcgtggatct cgcggtatca ttgcagcact
     7081 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     7141 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     7201 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     7261 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7321 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7381 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7441 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7501 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7561 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7621 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7681 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7741 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc
     7801 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7861 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7921 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7981 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     8041 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     8101 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
