LOCUS       Exported                6942 bp ds-DNA     circular SYN 06-JUN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6942)
  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..6942
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        complement(35..53)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(74..90)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    98..114
                     /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)."
     promoter        complement(122..152)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(476..1064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1235..2095)
                     /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"
     promoter        complement(2096..2200)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2203..2252
                     /label=oRH5055
     gene            2273..3629
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2273..2616
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2617..3426
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      3432..3629
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3641..3687)
                     /label=oRH5056
     promoter        3908..4283
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             4284..5999
                     /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"
     rep_origin      complement(6204..6659)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(join(6442..6942,1..78))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     6804..6820
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6830..6848
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(6857..6942,1..22)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     6874..6890
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6924..6940)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
       61 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      121 acaacatagg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgaggtaac
      181 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      241 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
      301 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      361 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      421 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      481 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      541 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      601 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      661 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      721 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      781 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      841 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      901 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      961 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1021 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1081 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1141 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1201 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1261 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1321 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     1381 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1441 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1501 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1561 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1621 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1681 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1741 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1801 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1861 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1921 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1981 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2041 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     2101 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     2161 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     2221 cacctgacgt cttgtttagc ttgcctcgtc cccgccgggt cacccggcca gcgacatgga
     2281 ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg
     2341 tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca tacattttga
     2401 tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa
     2461 cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa
     2521 aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg
     2581 atacagttct cacatcacat ccgaacataa acaaccatgg gtaaggaaaa gactcacgtt
     2641 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     2701 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
     2761 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     2821 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     2881 cctgatgatg catggttact caccactgcg atccccggca aaacagcatt ccaggtatta
     2941 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     3001 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     3061 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     3121 aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg
     3181 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     3241 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     3301 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     3361 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     3421 ttctaatcag tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt
     3481 ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc
     3541 gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca
     3601 atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag
     3661 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     3721 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     3781 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     3841 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagatct
     3901 gaattaattc ttgaataata cataactttt cttaaaagaa tcaaagacag ataaaattta
     3961 agagatatta aatattagtg agaagccgag aattttgtaa caccaacata acactgacat
     4021 ctttaacaac ttttaattat gatacatttc ttacgtcatg attgattatt acagctatgc
     4081 tgacaaatga ctcttgttgc atggctacga accgggtaat actaagtgat tgactcttgc
     4141 tgacctttta ttaagaacta aatggacaat attatggagc atttcatgta taaattggtg
     4201 cgtaaaatcg ttggatctct cttctaagta catcctacta taacaatcaa gaaaaacaag
     4261 aaaatcggac aaaacaatca agtatggatt ctagaacagt tggtatatta ggagggggac
     4321 aattgggacg tatgattgtt gaggcagcaa acaggctcaa cattaagacg gtaatactag
     4381 atgctgaaaa ttctcctgcc aaacaaataa gcaactccaa tgaccacgtt aatggctcct
     4441 tttccaatcc tcttgatatc gaaaaactag ctgaaaaatg tgatgtgcta acgattgaga
     4501 ttgagcatgt tgatgttcct acactaaaga atcttcaagt aaaacatccc aaattaaaaa
     4561 tttacccttc tccagaaaca atcagattga tacaagacaa atatattcaa aaagagcatt
     4621 taatcaaaaa tggtatagca gttacccaaa gtgttcctgt ggaacaagcc agtgagacgt
     4681 ccctattgaa tgttggaaga gatttgggtt ttccattcgt cttgaagtcg aggactttgg
     4741 catacgatgg aagaggtaac ttcgttgtaa agaataagga aatgattccg gaagctttgg
     4801 aagtactgaa ggatcgtcct ttgtacgccg aaaaatgggc accatttact aaagaattag
     4861 cagtcatgat tgtgaggtct gttaacggtt tagtgttttc ttacccaatt gtagagacta
     4921 tccacaagga caatatttgt gacttatgtt atgcgcctgc tagagttccg gactccgttc
     4981 aacttaaggc gaagttgttg gcagaaaatg caatcaaatc ttttcccggt tgtggtatat
     5041 ttggtgtgga aatgttctat ttagaaacag gggaattgct tattaacgaa attgccccaa
     5101 ggcctcacaa ctctggacat tataccattg atgcttgcgt cacttctcaa tttgaagctc
     5161 atttgagatc aatattggat ttgccaatgc caaagaattt cacatctttc tccaccatta
     5221 caacgaacgc cattatgcta aatgttcttg gagacaaaca tacaaaagat aaagagctag
     5281 aaacttgcga aagagcattg gcgactccag gttcctcagt gtacttatat ggaaaagagt
     5341 ctagacctaa cagaaaagta ggtcacataa atattattgc ctccagtatg gcggaatgtg
     5401 aacaaaggct gaactacatt acaggtagaa ctgatattcc aatcaaaatc tctgtcgctc
     5461 aaaagttgga cttggaagca atggtcaaac cattggttgg aatcatcatg ggatcagact
     5521 ctgacttgcc ggtaatgtct gccgcatgtg cggttttaaa agattttggc gttccatttg
     5581 aagtgacaat agtctctgct catagaactc cacataggat gtcagcatat gctatttccg
     5641 caagcaagcg tggaattaaa acaattatcg ctggagctgg tggggctgct cacttgccag
     5701 gtatggtggc tgcaatgaca ccacttcctg tcatcggtgt gcccgtaaaa ggttcttgtc
     5761 tagatggagt agattcttta cattcaattg tgcaaatgcc tagaggtgtt ccagtagcta
     5821 ccgtcgctat taataatagt acgaacgctg cgctgttggc tgtcagactg cttggcgctt
     5881 atgattcaag ttatacaacg aaaatggaac agtttttatt aaagcaagaa gaagaagttc
     5941 ttgtcaaagc acaaaagtta gaaactgtcg gttacgaagc ttatctagaa aacaagtaat
     6001 atataagttt attgatatac ttgtacagca aataattata aaatgatata cctatttttt
     6061 aggctttgtt atgattacat caaatgtgga cttcatacat agaaatcaac gcttacaggt
     6121 gtcctttttt aagaatttca tacataagat ctatgcggtg tgaaataccg cacagatgcg
     6181 taaggagaaa ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt
     6241 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     6301 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     6361 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     6421 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6481 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6541 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6601 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6661 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6721 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     6781 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
     6841 actatagggc gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga
     6901 tatcgaattc ctgcagcccg ggggatccac tagttctaga gc
//
