LOCUS       Exported                8144 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 8144)
  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..8144
                     /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..3982
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     terminator      4009..5000
                     /label=nmt1 terminator
     rep_origin      5080..5863
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6239..6343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6344..7204
                     /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      7375..7963
                     /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 gataaaggaa gaggaatcct
     3901 ggcatatcat caattgaata agttgaatta attatttcaa tctcattctc actttctgac
     3961 ttatagtcgc tttgttaaat catatgtcga ctctagagga tccccgggta aaaggaatgt
     4021 ctcccttgcc agtactgcta gggtttttct ttcaaactat ggaagcccat tcaagctgca
     4081 tattacgatt ttgtttttcg cttttagaaa gtggtttaga tgagataata gaaaaattct
     4141 tgatctccga caacgagtac ttttattttt tttgctaatc actttactca atattagctc
     4201 gaaatcgtag aaacgtagac gggtgcggga taccgagtgg tgtagttaag aatttttata
     4261 aaccacgtgg cccaaaaata tgaacccaaa acgtttatac atgagtatac tttaagaagg
     4321 ctatacccct tcgtgttaga tgtagtttta gctacccaac ccgagtctat gagcttgact
     4381 tcagatgtag aaggcattaa atcgttttga atattaatta aaaaacgatg aaaattaaat
     4441 atttaaaagc aatcatacgc tgaaaattta gtgctgtggc taatccttca acatggaaat
     4501 gccataaaag tgactttgac aaaaaaaaaa gtatatacag gtagtaaact catctacttc
     4561 attgactttg tttacagcat gtggaaggag gaatatttat tgctaaatcg tagtttaaca
     4621 ttcaataagt aatactattg aaattcgaca agattggccg catggatgaa aaagaggcat
     4681 tttgctttgg gagaattagt tcaaattaga actgaaaaaa aaaactttac gaggcaaaaa
     4741 tgtcggattg agatcgtaaa agttcgctcg tcgtcttttg ctttgtgatt gttttcatgg
     4801 atacatcttg ctggatattt aaattttagt actatgtata agatattcta taaatgtttt
     4861 atcacccaaa cctgttagcg ccttcttaat tctattcaat ctggcttttg ctctgagact
     4921 acttcttgga ctttcactac ttgttagtta tacggaattt gtgtaattag aagtgaaata
     4981 atcctttcta ttagtaatgc gagctgcaat tcgagtctaa ctccttaacc actcggacat
     5041 agtgacttat ctgacactca ttgtaaatta atatatatat aggcattttg tttagttaaa
     5101 ggtacttaag taattagtat aaacgaacca attttataat caggaagtta agtgaatggt
     5161 agcacatgtc gtaaaaattg tgaattttta ttgaataata ttttaaatac aagcctttct
     5221 agactaggta tactcataaa catatatgag caaaaggata gaggagatta cattgcatct
     5281 tctacaaatt tatttattgc cctttactga aaaattaaat aatgagtact aaatgataaa
     5341 aagcgctcag tacaagaaag attgcaaaaa tattgcattc ttcatgaatt aaagttgcat
     5401 ataaggcata ttgaaagtaa tagtactaaa acagcagtta gcgaaaatta atagaattat
     5461 attcgcaaga caattgtgac attaaattaa aaaattgtag aatttttact atcctcttta
     5521 acgccatgag cctttataaa aaggttaaat tagttttaac attctttttt tgagtaagag
     5581 tttacaattt atcaaaacct gtgttattat attcattagt ttcaatttat tagcatctag
     5641 agaaaaatca attggcagtt acattgttgg aatttatgaa gaaaaagaat ctacaacgga
     5701 gaataagttg ctgatcgctt tccctaaaat tgtatatttt gctgagctta ttttgacatt
     5761 tcgttgaagt tttctctaat tcgcattcat tttaagtaaa acaatgagaa ataaaattac
     5821 aaaaaataca aattaaaata caatttttag ctataattat agacgatgcc cttgtatccc
     5881 attctgtctc gcttgcccct actttttatc ttttatatac cataatgaac gctgccgcta
     5941 ctaaccatac cccgatttta catttcggac tcccaaggac gtacaaaata gaaaactata
     6001 gaaaaaaata atcagaaaat agcatgtcat ctctttgtaa aacgcgtttg caagaagaaa
     6061 ggaaacaatg gagaagagat catccatttg tatgtaaatt ttagtaaact tgaagaaatc
     6121 actaacaact tctcttactt agggattcta tgcaaaacct tgtaaatcat ctgatggagg
     6181 actcgattta atgaattgga aggttggatt tccaggtggc acttttcggg gaaatgtgcg
     6241 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6301 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6361 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6421 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6481 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6541 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6601 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6661 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6721 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6781 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6841 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6901 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6961 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7021 ctggtttatt gctgataaat ctggagccgg tgagcgtgga tctcgcggta tcattgcagc
     7081 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7141 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7201 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7261 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7321 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7381 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7441 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7501 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     7561 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7621 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7681 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7741 cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa
     7801 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7861 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7921 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7981 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     8041 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     8101 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
