LOCUS       Exported                7641 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast marker swap plasmid for converting HIS3 to ADE2.
ACCESSION   .
VERSION     .
KEYWORDS    M2371 his3::ADE2 Disruptor Converter
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7641)
  AUTHORS   Voth WP, Jiang YW, Stillman DJ
  TITLE     New 'marker swap' plasmids for converting selectable markers on 
            budding yeast gene disruptions and plasmids.
  JOURNAL   Yeast. 2003 Aug;20(11):985-93.
  PUBMED    12898713
REFERENCE   2  (bases 1 to 7641)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7641
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        711..898
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     1113..1129
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1113..1129
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1543..1554
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             complement(1853..3568)
                     /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(3569..3944)
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     primer_bind     complement(5420..5436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5420..5436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5433..5455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5444..5460
                     /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(5468..5498)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5513..5534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5651..5668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5822..6410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5902..5921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6581..7441)
                     /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"
     primer_bind     7204..7223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7442..7546)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7614..7632
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 actctagagg atccgctgca cggtcctgtt ccctagcatg tacgtgagcg tatttccttt
      481 taaaccacga cgctttgtct tcattcaacg tttcccattg tttttttcta ctattgcttt
      541 gctgtgggaa aaacttatcg aaagatgacg actttttctt aattctcgtt ttaagagctt
      601 ggtgagcgct aggagtcact gccaggtatc gtttgaacac ggcattagtc agggaagtca
      661 taacacagtc ctttcccgca attttctttt tctattactc ttggcctcct ctagtacact
      721 ctatattttt ttatgcctcg gtaatgattt tcattttttt ttttccacct agcggatgac
      781 tctttttttt tcttagcgat tggcattatc acataatgaa ttatacatta tataaagtaa
      841 tgtgatttct tcgaagaata tactaaaaaa tgagcaggca agataaacga aggcaaagat
      901 gacagagcag aaagccctag taaagcgtat tacaaatgaa accaagattc agattgcgat
      961 ctctttaaag ggtggtcccc tagcgataga gcactcgatc ttcccagaaa aagaggcaga
     1021 agcagtagca gaacaggcca cacaatcgca agtgattaac gtccacacag gtatagggtt
     1081 tctggaccat atgatacatg ctctggggcc gctctagaac tagtggatcc gggatctcaa
     1141 cgttgttact ggtgtgttga agcgatactt aagaaagctc cccaacccta tctttacctt
     1201 tcaaatatac gagccgttga tgaggttagt taaatctaaa aaaatgatgg aaaacttgcc
     1261 atttgttgga ggaaagttgt ccttagaagc aaagaattcg gacacttata tgtcgagcaa
     1321 gagcgcttta aaaaacatat tggaagacct tccaagggaa cattataggg tgttaagagt
     1381 actgagtgaa catatagaaa aggttacacg gtacagtcac tggaatcgaa tgacgcttta
     1441 taatttggct ttagtttttg ctccaggttt gattcgtgat ttcagtggag aaaaggatat
     1501 tattgatatg aaagaaagaa actatattgt cgcatttata tttgggaact acaaagatat
     1561 cctgacgtag cgctatcctc ggttctgcat tgagccgcct tatatgaact gtatcgaaac
     1621 gttatttttt taatcgcaga cttaagcagg taattattcc ttgcttcttg ttactggata
     1681 tgtatgtatg tataataagt gatcttatgt atgaaattct taaaaaagga cacctgtaag
     1741 cgttgatttc tatgtatgaa gtccacattt gatgtaatca taacaaagcc taaaaaatag
     1801 gtatatcatt ttataattat ttgctgtaca agtatatcaa taaacttata tattacttgt
     1861 tttctagata agcttcgtaa ccgacagttt ctaacttttg tgctttgaca agaacttctt
     1921 cttcttgctt taataaaaac tgttccattt tcgttgtata acttgaatca taagcgccaa
     1981 gcagtctgac agccaacagc gcagcgttcg tactattatt aatagcgacg gtagctactg
     2041 gaacacctct aggcatttgc acaattgaat gtaaagaatc tactccatct agacaagaac
     2101 cttttacggg cacaccgatg acaggaagtg gtgtcattgc agccaccata cctggcaagt
     2161 gagcagcccc accagctcca gcgataattg ttttaattcc acgcttgctt gcggaaatag
     2221 catatgctga catcctatgt ggagttctat gagcagagac tattgtcact tcaaatggaa
     2281 cgccaaaatc ttttaaaacc gcacatgcgg cagacattac cggcaagtca gagtctgatc
     2341 ccatgatgat tccaaccaat ggtttgacca ttgcttccaa gtccaacttt tgagcgacag
     2401 agattttgat tggaatatca gttctacctg taatgtagtt cagcctttgt tcacattccg
     2461 ccatactgga ggcaataata tttatgtgac ctacttttct gttaggtcta gactcttttc
     2521 catataagta cactgaggaa cctggagtcg ccaatgctct ttcgcaagtt tctagctctt
     2581 tatcttttgt atgtttgtct ccaagaacat ttagcataat ggcgttcgtt gtaatggtgg
     2641 agaaagatgt gaaattcttt ggcattggca aatccaatat tgatctcaaa tgagcttcaa
     2701 attgagaagt gacgcaagca tcaatggtat aatgtccaga gttgtgaggc cttggggcaa
     2761 tttcgttaat aagcaattcc cctgtttcta aatagaacat ttccacacca aatataccac
     2821 aaccgggaaa agatttgatt gcattttctg ccaacaactt cgccttaagt tgaacggagt
     2881 ccggaactct agcaggcgca taacataagt cacaaatatt gtccttgtgg atagtctcta
     2941 caattgggta agaaaacact aaaccgttaa cagatctcac aatcatgact gctaattctt
     3001 tagtaaatgg tgcccatttt tcggcgtaca aaggacgatc cttcagtact tccaaagctt
     3061 ccggaatcat ttccttattc tttacaacga agttacctct tccatcgtat gccaaagtcc
     3121 tcgacttcaa gacgaatgga aaacccaaat ctcttccaac attcaatagg gacgtctcac
     3181 tggcttgttc cacaggaaca ctttgggtaa ctgctatacc atttttgatt aaatgctctt
     3241 tttgaatata tttgtcttgt atcaatctga ttgtttctgg agaagggtaa atttttaatt
     3301 tgggatgttt tacttgaaga ttctttagtg taggaacatc aacatgctca atctcaatcg
     3361 ttagcacatc acatttttca gctagttttt cgatatcaag aggattggaa aaggagccat
     3421 taacgtggtc attggagttg cttatttgtt tggcaggaga attttcagca tctagtatta
     3481 ccgtcttaat gttgagcctg tttgctgcct caacaatcat acgtcccaat tgtccccctc
     3541 ctaatatacc aactgttcta gaatccatac ttgattgttt tgtccgattt tcttgttttt
     3601 cttgattgtt atagtaggat gtacttagaa gagagatcca acgattttac gcaccaattt
     3661 atacatgaaa tgctccataa tattgtccat ttagttctta ataaaaggtc agcaagagtc
     3721 aatcacttag tattacccgg ttcgtagcca tgcaacaaga gtcatttgtc agcatagctg
     3781 taataatcaa tcatgacgta agaaatgtat cataattaaa agttgttaaa gatgtcagtg
     3841 ttatgttggt gttacaaaat tctcggcttc tcactaatat ttaatatctc ttaaatttta
     3901 tctgtctttg attcttttaa gaaaagttat gtattattca agaaaaagtc aattccgcat
     3961 caaaaggtaa aatttatata aactgcttta aaatttcatg aaactaggca acttttcgaa
     4021 atgatctttt tcgagcatga agtttctttt ataataacct ggtcaaaagc tttcaatata
     4081 taatacattt ggtatttacg gaaatgagat gatatactgg tagtgcgcta gtcaagttct
     4141 aattaaatta aaatgcaaaa tagtaatgtc agatggacac attgggcgaa atagctaacc
     4201 cataagcatg aatcctgtaa cttatgtaat acgttaatca cgatacggcg ttactagtct
     4261 tccttttgag tggtttctaa tgtagattct tgttgttcgt taatttgctt caagccgttt
     4321 agcattcctg attcgtttga agatagagga acagatgcgc cagcgaaaga tgatgatgca
     4381 atggtagagg aagtgttggt cgaatctccc tttttggaaa taaagtgtcg ttttgattct
     4441 agataggctt ctcgcaattg attgtggaga tacttcttta aaagggccat acctatggga
     4501 tgacattcta acctatagag taatttcgta atagcatagc agatttttga cgcccttctt
     4561 tctagtagag gagaagccct tagcttgggt gagatgactg aggatgcgtt tgtaaattcg
     4621 ggataattgg aatttccagt accctgccca ttacctccca gaatccttat attagggcgt
     4681 gtactcagtt gtaaaaactg tctcaatgaa gaagattgag caattctggt caaatacgtt
     4741 cttctcctgg acagagattt tgtgaagggc aaatcctgcc cactttcttt tgtaataaat
     4801 gtgaggaaaa acgccaagat atcaaaactg taaatttgag taaaaagatc ccggatcccc
     4861 cgggctgcat cattctatac gtgtcattct gaacgaggcg cgctttcctt ttttcttttt
     4921 gctttttctt tttttttctc ttgaactcga gaaaaaaaat ataaaagaga tggaggaacg
     4981 ggaaaaagtt agttgtggtg ataggtggca agtggtattc cgtaagaaca acaagaaaag
     5041 catttcatat tatggctgaa ctgagcgaac aagtgcaaaa tttaagcatc aacgacaaca
     5101 acgagaatgg ttatgttcct cctcacttaa gaggaaaacc aagaagtgcc agaaataaca
     5161 gtagcaacta caataacaac aacggcggct acaacggtgg ccgtggcggt ggcagcttct
     5221 ttagcaacaa ccgtcgtggt ggttacggca acggtggttt cttcggtgga aacaacggtg
     5281 gcagcagatc taacggccgt tctggtggta gatggatcga tggcaaacat gtcccagctc
     5341 caagaaacga aaaggccgag atcgccatat ttggtgtccc cgaggatccc cgggtaccga
     5401 gctcgaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5461 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5521 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5581 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5641 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5701 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5761 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5821 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5881 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5941 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6001 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6061 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6181 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6241 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6301 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6361 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6421 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6481 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6541 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6601 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6661 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6721 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6781 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6841 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6901 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6961 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7021 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7081 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7141 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7201 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7261 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7321 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7381 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7441 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7501 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7561 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     7621 gtatcacgag gccctttcgt c
//
