LOCUS       Exported                7426 bp ds-DNA     circular SYN 26-JAN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7426)
  AUTHORS   Hampton lab
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2019 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7426
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     3..19
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(53..69)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(95..113)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(123..139)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      284..739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(944..2659)
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     promoter        complement(2660..3035)
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             complement(3548..4642)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVAGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4655..5059)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        5227..5331
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5332..6192
                     /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      6363..6951
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        7275..7305
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7313..7329
                     /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     7337..7353
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(7349..7426,1..501)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     promoter        7374..7392
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(7405..7426,1..86)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
ORIGIN
        1 gctctagaac tagtggatcc cccgggctgc aggaattcga tatcaagctt atcgataccg
       61 tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc
      121 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
      181 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
      241 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc
      301 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      361 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      421 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      481 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      541 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      601 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
      661 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
      721 atattaacgt ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
      781 cacaccgcat agatcttatg tatgaaattc ttaaaaaagg acacctgtaa gcgttgattt
      841 ctatgtatga agtccacatt tgatgtaatc ataacaaagc ctaaaaaata ggtatatcat
      901 tttataatta tttgctgtac aagtatatca ataaacttat atattacttg ttttctagat
      961 aagcttcgta accgacagtt tctaactttt gtgctttgac aagaacttct tcttcttgct
     1021 ttaataaaaa ctgttccatt ttcgttgtat aacttgaatc ataagcgcca agcagtctga
     1081 cagccaacag cgcagcgttc gtactattat taatagcgac ggtagctact ggaacacctc
     1141 taggcatttg cacaattgaa tgtaaagaat ctactccatc tagacaagaa ccttttacgg
     1201 gcacaccgat gacaggaagt ggtgtcattg cagccaccat acctggcaag tgagcagccc
     1261 caccagctcc agcgataatt gttttaattc cacgcttgct tgcggaaata gcatatgctg
     1321 acatcctatg tggagttcta tgagcagaga ctattgtcac ttcaaatgga acgccaaaat
     1381 cttttaaaac cgcacatgcg gcagacatta ccggcaagtc agagtctgat cccatgatga
     1441 ttccaaccaa tggtttgacc attgcttcca agtccaactt ttgagcgaca gagattttga
     1501 ttggaatatc agttctacct gtaatgtagt tcagcctttg ttcacattcc gccatactgg
     1561 aggcaataat atttatgtga cctacttttc tgttaggtct agactctttt ccatataagt
     1621 acactgagga acctggagtc gccaatgctc tttcgcaagt ttctagctct ttatcttttg
     1681 tatgtttgtc tccaagaaca tttagcataa tggcgttcgt tgtaatggtg gagaaagatg
     1741 tgaaattctt tggcattggc aaatccaata ttgatctcaa atgagcttca aattgagaag
     1801 tgacgcaagc atcaatggta taatgtccag agttgtgagg ccttggggca atttcgttaa
     1861 taagcaattc ccctgtttct aaatagaaca tttccacacc aaatatacca caaccgggaa
     1921 aagatttgat tgcattttct gccaacaact tcgccttaag ttgaacggag tccggaactc
     1981 tagcaggcgc ataacataag tcacaaatat tgtccttgtg gatagtctct acaattgggt
     2041 aagaaaacac taaaccgtta acagacctca caatcatgac tgctaattct ttagtaaatg
     2101 gtgcccattt ttcggcgtac aaaggacgat ccttcagtac ttccaaagct tccggaatca
     2161 tttccttatt ctttacaacg aagttacctc ttccatcgta tgccaaagtc ctcgacttca
     2221 agacgaatgg aaaacccaaa tctcttccaa cattcaatag ggacgtctca ctggcttgtt
     2281 ccacaggaac actttgggta actgctatac catttttgat taaatgctct ttttgaatat
     2341 atttgtcttg tatcaatctg attgtttctg gagaagggta aatttttaat ttgggatgtt
     2401 ttacttgaag attctttagt gtaggaacat caacatgctc aatctcaatc gttagcacat
     2461 cacatttttc agctagtttt tcgatatcaa gaggattgga aaaggagcca ttaacgtggt
     2521 cattggagtt gcttatttgt ttggcaggag aattttcagc atctagtatt accgtcttaa
     2581 tgttgagcct gtttgctgcc tcaacaatca tacgtcccaa ttgtccccct cctaatatac
     2641 caactgttct agaatccata cttgattgtt ttgtccgatt ttcttgtttt tcttgattgt
     2701 tatagtagga tgtacttaga agagagatcc aacgatttta cgcaccaatt tatacatgaa
     2761 atgctccata atattgtcca tttagttctt aataaaaggt cagcaagagt caatcactta
     2821 gtattacccg gttcgtagcc atgcaacaag agtcatttgt cagcatagct gtaataatca
     2881 atcatgacgt aagaaatgta tcataattaa aagttgttaa agatgtcagt gttatgttgg
     2941 tgttacaaaa ttctcggctt ctcactaata tttaatatct cttaaatttt atctgtcttt
     3001 gattctttta agaaaagtta tgtattattc aagaattaat tcagatctat ggtgcactct
     3061 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3121 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3181 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3241 gggcctcgtg atacgcctat ttttataggt taatgtcatg atctttttgt gtggtgccct
     3301 cctccttgtc aatattaatg ttaaagtgca attctttttc cttatcacgt tgagccatta
     3361 gtatcaattt gcttacctgt attcctttac tatcctcctt tttctccttc ttgataaatg
     3421 tatgtagatt gcgtatatag tttcgtctac cctatgaaca tattccattt tgtaatttcg
     3481 tgtcgtttct attatgaatt tcatttataa agtttatgta caaatatcat aaaaaaagag
     3541 aatcttttta agcaaggatt ttcttaactt cttcggcgac agcatcaccg acttcggtgg
     3601 tactgttgga accacctaaa tcaccagttc tgatacctgc atccaaaacc tttttaactg
     3661 catcttcaat ggccttacct tcttcaggca agttcaatga caatttcaac atcattgcag
     3721 cagacaagat agtggcgata gggtcaacct tattctttgg caaatctgga gcagaaccgt
     3781 ggcatggttc gtacaaacca aatgcggtgt tcttgtctgg caaagaggcc aaggacgcag
     3841 atggcaacaa acccaaggaa cctgggataa cggaggcttc atcggagatg atatcaccaa
     3901 acatgttgct ggtgattata ataccattta ggtgggttgg gttcttaact aggatcatgg
     3961 cggcagaatc aatcaattga tgttgaacct tcaatgtagg gaattcgttc ttgatggttt
     4021 cctccacagt ttttctccat aatcttgaag aggccaaaag attagcttta tccaaggacc
     4081 aaataggcaa tggtggctca tgttgtaggg ccatgaaagc ggccattctt gtgattcttt
     4141 gcacttctgg aacggtgtat tgttcactat cccaagcgac accatcacca tcgtcttcct
     4201 ttctcttacc aaagtaaata cctcccacta attctctgac aacaacgaag tcagtacctt
     4261 tagcaaattg tggcttgatt ggagataagt ctaaaagaga gtcggatgca aagttacatg
     4321 gtcttaagtt ggcgtacaat tgaagttctt tacggatttt tagtaaacct tgttcaggtc
     4381 taacactacc ggtaccccat ttaggaccag ccacagcacc taacaaaacg gcatcaacct
     4441 tcttggaggc ttccagcgcc tcatctggaa gtgggacacc tgtagcatcg atagcagcac
     4501 caccaattaa atgattttcg aaatcgaact tgacattgga acgaacatca gaaatagctt
     4561 taagaacctt aatggcttcg gctgtgattt cttgaccaac gtggtcacct ggcaaaacga
     4621 cgatcttctt aggggcagac ataggggcag acattagaat ggtatatcct tgaaatatat
     4681 atatatattg ctgaaatgta aaaggtaaga aaagttagaa agtaagacga ttgctaacca
     4741 cctattggaa aaaacaatag gtccttaaat aatattgtca acttcaagta ttgtgatgca
     4801 agcatttagt catgaacgct tctctattct atatgaaaag ccggttccgg cctctcacct
     4861 ttcctttttc tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat
     4921 taacaaaaaa tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc
     4981 gttatgttga ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata
     5041 cctgagtatt cccacagtta actgcggtca agatatttct tgaatcaggc gccttagacc
     5101 gctcggccaa acaaccaatt acttgttgag aaatagagta taattatcct ataaatataa
     5161 cgtttttgaa cacacatgaa caaggaagta caggagacgt caggtggcac ttttcgggga
     5221 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5281 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5341 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5401 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5461 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5521 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5581 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5641 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5701 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5761 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5821 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5881 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5941 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6001 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6061 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6121 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6181 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6241 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6301 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6361 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6421 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6481 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     6541 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6601 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6661 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6721 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6781 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6841 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6901 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6961 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7021 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7081 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7141 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7201 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta cctcactcat
     7261 taggcacccc aggctttaca ctttatgctt ccggctccta tgttgtgtgg aattgtgagc
     7321 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     7381 cctcactaaa gggaacaaaa gctggagctc caccgcggtg gcggcc
//
