LOCUS       Exported                5896 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 5896)
  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..5896
                     /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(1102..2540)
                     /label=S. pombe his3
     CDS             complement(1277..2431)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his3"
                     /product="imidazole acetol phosphate transaminase, required
                     for histidine biosynthesis"
                     /label=his3 CDS
                     /translation="MFDLNTCLRKNILELQPYRCARDDFSEGVLLDANECAYGSVISVD
                     GVEFNRYPDPRQIEVKQRLCDLRNKELSITKPLTPDNICMGVGSDEIIDSLIRISCIPG
                     KDKILMCPPSYGMYTVSAKINDVEVVKVLLEPDFNLNVDAICETLSKDSAIKVFFACSP
                     GNPTAKALKLEDIKKILEHPTWNGIVVVDEAYIDFSAPDMSALTLVNEYPNLAVCQTLS
                     KSFGLAGIRIGFCLTSKPIATIMNSLKAPYNISEPTSRLALDALSPQSIDKMHTYRDAI
                     IQQRVRLCKELTTIKGMGKIIGGYDANFILIQVLDRPEGGKPSNDAAKYLYLQMATMHK
                     VVVRFRGTEPLCEGALRITVGTEEENTILLKTIKLVLQEYYTKK"
     misc_feature    1740..1744
                     /label=one intron deleted here
     misc_feature    2329..2386
                     /label=two introns removed here
     misc_feature    complement(2432..2540)
                     /label=5' UTR
     rep_origin      2832..3615
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        3991..4095
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4096..4956
                     /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      5127..5715
                     /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 tttaaattag cgtaactcct ttacaaaatt aaaaataaca tcttttcatt
     1081 gcttatcgaa ctatacggaa acacaccaaa aaggaagaaa acgtatttta ataaagaaag
     1141 aaggaatgca agcgcaatta gcgaacacaa aatatattgg gtacatcaga aaagaaatgt
     1201 gacgagtccg gaaggggaaa ggaaagagtt ataatttgtc taattgaaac gttctaatag
     1261 gccagaaaag gaaacctcat ttttttgtat agtattcctg caatacaagt ttaatagttt
     1321 ttagtaatat cgtattctct tcttctgtgc caacagtgat acgcaatgct ccttcgcaaa
     1381 gaggctctgt acctctaaat ctaacaacga ctttgtgcat cgtggccatt tgtaaataaa
     1441 gatattttgc ggcatcgtta ctaggcttac ctccttctgg tctatctaaa acctgaatga
     1501 ggataaaatt tgcatcataa ccacctataa tcttgcccat tcctttaatg gtagtaagct
     1561 ctttgcaaag acgaactctt tgctgtataa tagcatctct atatgtatgc attttatcaa
     1621 tcgactgagg ggataaagca tccaaagcta aacgagaggt gggttcgcta atgttataag
     1681 gagcctttaa tgagttcata atggtagcaa taggtttact tgtcaaacaa aatccgatac
     1741 gaatacctgc tagaccgaaa gatttcgaaa gggtttggca aacagcaagg tttggatatt
     1801 cattgacaag agttaaggca gacatatcgg gagcagagaa atcaatatag gcctcatcga
     1861 cgacaacgat tccattccat gtagggtgct ccaaaatctt cttaatatcc tccagtttaa
     1921 gagccttagc tgttggatta ccaggggaac aagcaaaaaa gactttgatg gcactatctt
     1981 tagaaagtgt ttcgcaaata gcatcgacgt tcaaattgaa atctggttca aggagaacct
     2041 taacaacctc aacatcgttg atttttgcag acaccgtata cattccgtac gaaggtggac
     2101 acatcaaaat cttatctttt ccaggaatac aagaaatgcg aattaaggag tcaataattt
     2161 catcgctgcc aacacccatg caaatgttat ctggtgttag tggcttggta atggaaagct
     2221 ctttgtttct caaatcacat aatctttgct ttacttcgat ctgtctagga tcgggataac
     2281 gattaaattc cacaccgtcc acggaaataa ccgaaccata agcacattca ttggcatcaa
     2341 gaaggacacc ttcagagaag tcatctctcg cacagcgata aggctgaagt tctaagatat
     2401 ttttacgaag acaagtattc aaatcaaaca tcgtatctct ttcgatgtgt ttttgttaaa
     2461 gtaagaagaa agcgatcgat gagattaatt aataattaac ctaagatgca ggtgatatct
     2521 ataaatatat agtgtgtagc aatgtgagtc tttaattcat tacaacgtgt atcaccgtgt
     2581 cattatatcc aataaattca gaaacctgaa gaggaaagag aaataaagct cgatatgtct
     2641 ttcatttcca aatacaatta taagtgcaat agtaaagaaa acgttgaaag cttgataaaa
     2701 atttcgttac tatactaatg ctgtaaagaa aatgcaccat gtcgatcgac tctagaggaa
     2761 attcgagtct aactccttaa ccactcggac atagtgactt atctgacact cattgtaaat
     2821 taatatatat ataggcattt tgtttagtta aaggtactta agtaattagt ataaacgaac
     2881 caattttata atcaggaagt taagtgaatg gtagcacatg tcgtaaaaat tgtgaatttt
     2941 tattgaataa tattttaaat acaagccttt ctagactagg tatactcata aacatatatg
     3001 agcaaaagga tagaggagat tacattgcat cttctacaaa tttatttatt gccctttact
     3061 gaaaaattaa ataatgagta ctaaatgata aaaagcgctc agtacaagaa agattgcaaa
     3121 aatattgcat tcttcatgaa ttaaagttgc atataaggca tattgaaagt aatagtacta
     3181 aaacagcagt tagcgaaaat taatagaatt atattcgcaa gacaattgtg acattaaatt
     3241 aaaaaattgt agaattttta ctatcctctt taacgccatg agcctttata aaaaggttaa
     3301 attagtttta acattctttt tttgagtaag agtttacaat ttatcaaaac ctgtgttatt
     3361 atattcatta gtttcaattt attagcatct agagaaaaat caattggcag ttacattgtt
     3421 ggaatttatg aagaaaaaga atctacaacg gagaataagt tgctgatcgc tttccctaaa
     3481 attgtatatt ttgctgagct tattttgaca tttcgttgaa gttttctcta attcgcattc
     3541 attttaagta aaacaatgag aaataaaatt acaaaaaata caaattaaaa tacaattttt
     3601 agctataatt atagacgatg cccttgtatc ccattctgtc tcgcttgccc ctacttttta
     3661 tcttttatat accataatga acgctgccgc tactaaccat accccgattt tacatttcgg
     3721 actcccaagg acgtacaaaa tagaaaacta tagaaaaaaa taatcagaaa atagcatgtc
     3781 atctctttgt aaaacgcgtt tgcaagaaga aaggaaacaa tggagaagag atcatccatt
     3841 tgtatgtaaa ttttagtaaa cttgaagaaa tcactaacaa cttctcttac ttagggattc
     3901 tatgcaaaac cttgtaaatc atctgatgga ggactcgatt taatgaattg gaaggttgga
     3961 tttccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4021 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4081 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4141 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4201 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4261 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4321 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4381 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4441 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4501 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4561 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4621 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4681 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4741 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4801 ggtgagcgtg gatctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4861 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4921 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4981 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5041 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5101 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5161 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5221 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     5281 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5341 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5401 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5461 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat
     5521 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5581 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5641 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5701 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5761 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5821 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5881 agcgaggaag cggaag
//
