LOCUS       Exported                7161 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 7161)
  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..7161
                     /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..3848
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        2273..2616
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2617..3645
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      3651..3848
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3860..3906)
                     /label=oRH5056
     promoter        4127..4502
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             4503..6218
                     /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(6423..6878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(join(6661..7161,1..78))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     7023..7039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7042..7068
                     /label=Recomb OH 1
     promoter        7049..7067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(7076..7161,1..22)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     7093..7109
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7143..7159)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(7159..7161,1..7))
                     /label=Recomb OH 2
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 gtaaaaagcc tgaactcacc
     2641 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
     2701 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
     2761 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     2821 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg
     2881 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
     2941 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt
     3001 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
     3061 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     3121 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     3181 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     3241 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     3301 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     3361 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc
     3421 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     3481 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
     3541 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
     3601 agtggaaacc gacgccccag cactcgtccg agggcaaagg aataatcagt actgacaata
     3661 aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc
     3721 tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag
     3781 atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg
     3841 ctatactgct gtcgattcga tactaacgcc gccatccaga tcatgacatt aacctataaa
     3901 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3961 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4021 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     4081 gcatcagagc agattgtact gagagtgcac catagatctg aattaattct tgaataatac
     4141 ataacttttc ttaaaagaat caaagacaga taaaatttaa gagatattaa atattagtga
     4201 gaagccgaga attttgtaac accaacataa cactgacatc tttaacaact tttaattatg
     4261 atacatttct tacgtcatga ttgattatta cagctatgct gacaaatgac tcttgttgca
     4321 tggctacgaa ccgggtaata ctaagtgatt gactcttgct gaccttttat taagaactaa
     4381 atggacaata ttatggagca tttcatgtat aaattggtgc gtaaaatcgt tggatctctc
     4441 ttctaagtac atcctactat aacaatcaag aaaaacaaga aaatcggaca aaacaatcaa
     4501 gtatggattc tagaacagtt ggtatattag gagggggaca attgggacgt atgattgttg
     4561 aggcagcaaa caggctcaac attaagacgg taatactaga tgctgaaaat tctcctgcca
     4621 aacaaataag caactccaat gaccacgtta atggctcctt ttccaatcct cttgatatcg
     4681 aaaaactagc tgaaaaatgt gatgtgctaa cgattgagat tgagcatgtt gatgttccta
     4741 cactaaagaa tcttcaagta aaacatccca aattaaaaat ttacccttct ccagaaacaa
     4801 tcagattgat acaagacaaa tatattcaaa aagagcattt aatcaaaaat ggtatagcag
     4861 ttacccaaag tgttcctgtg gaacaagcca gtgagacgtc cctattgaat gttggaagag
     4921 atttgggttt tccattcgtc ttgaagtcga ggactttggc atacgatgga agaggtaact
     4981 tcgttgtaaa gaataaggaa atgattccgg aagctttgga agtactgaag gatcgtcctt
     5041 tgtacgccga aaaatgggca ccatttacta aagaattagc agtcatgatt gtgaggtctg
     5101 ttaacggttt agtgttttct tacccaattg tagagactat ccacaaggac aatatttgtg
     5161 acttatgtta tgcgcctgct agagttccgg actccgttca acttaaggcg aagttgttgg
     5221 cagaaaatgc aatcaaatct tttcccggtt gtggtatatt tggtgtggaa atgttctatt
     5281 tagaaacagg ggaattgctt attaacgaaa ttgccccaag gcctcacaac tctggacatt
     5341 ataccattga tgcttgcgtc acttctcaat ttgaagctca tttgagatca atattggatt
     5401 tgccaatgcc aaagaatttc acatctttct ccaccattac aacgaacgcc attatgctaa
     5461 atgttcttgg agacaaacat acaaaagata aagagctaga aacttgcgaa agagcattgg
     5521 cgactccagg ttcctcagtg tacttatatg gaaaagagtc tagacctaac agaaaagtag
     5581 gtcacataaa tattattgcc tccagtatgg cggaatgtga acaaaggctg aactacatta
     5641 caggtagaac tgatattcca atcaaaatct ctgtcgctca aaagttggac ttggaagcaa
     5701 tggtcaaacc attggttgga atcatcatgg gatcagactc tgacttgccg gtaatgtctg
     5761 ccgcatgtgc ggttttaaaa gattttggcg ttccatttga agtgacaata gtctctgctc
     5821 atagaactcc acataggatg tcagcatatg ctatttccgc aagcaagcgt ggaattaaaa
     5881 caattatcgc tggagctggt ggggctgctc acttgccagg tatggtggct gcaatgacac
     5941 cacttcctgt catcggtgtg cccgtaaaag gttcttgtct agatggagta gattctttac
     6001 attcaattgt gcaaatgcct agaggtgttc cagtagctac cgtcgctatt aataatagta
     6061 cgaacgctgc gctgttggct gtcagactgc ttggcgctta tgattcaagt tatacaacga
     6121 aaatggaaca gtttttatta aagcaagaag aagaagttct tgtcaaagca caaaagttag
     6181 aaactgtcgg ttacgaagct tatctagaaa acaagtaata tataagttta ttgatatact
     6241 tgtacagcaa ataattataa aatgatatac ctatttttta ggctttgtta tgattacatc
     6301 aaatgtggac ttcatacata gaaatcaacg cttacaggtg tcctttttta agaatttcat
     6361 acataagatc tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     6421 ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     6481 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     6541 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     6601 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     6661 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     6721 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     6781 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     6841 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc cattcaggct
     6901 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6961 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     7021 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
     7081 cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagcccgg
     7141 gggatccact agttctagag c
//
