LOCUS       Exported                6927 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast marker swap plasmid for converting HIS3 to KanMX3.
ACCESSION   .
VERSION     .
KEYWORDS    M3929 his3::KanMX3 Disruptor Converter
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6927)
  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 6927)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6927
                     /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"
     gene            1725..3081
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1725..2068
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2069..2878
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2136..2155)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2884..3081
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(4206..4393)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(4706..4722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4706..4722)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4719..4741)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4730..4746
                     /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(4754..4784)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4799..4820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4937..4954)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5108..5696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5188..5207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5867..6727)
                     /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     6490..6509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6728..6832)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6900..6918
                     /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 atcctcgggg acaccaaata tggcgatctc ggccttttcg tttcttggag
      481 ctgggacatg tttgccatcg atccatctac caccagaacg gccgttagat ctgctgccac
      541 cgttgtttcc accgaagaaa ccaccgttgc cgtaaccacc acgacggttg ttgctaaaga
      601 agctgccacc gccacggcca ccgttgtagc cgccgttgtt gttattgtag ttgctactgt
      661 tatttctggc acttcttggt tttcctctta agtgaggagg aacataacca ttctcgttgt
      721 tgtcgttgat gcttaaattt tgcacttgtt cgctcagttc agccataata tgaaatgctt
      781 ttcttgttgt tcttacggaa taccacttgc cacctatcac cacaactaac tttttcccgt
      841 tcctccatct cttttatatt ttttttctcg agttcaagag aaaaaaaaag aaaaagcaaa
      901 aagaaaaaag gaaagcgcgc ctcgttcaga atgacacgta tagaatgatg cattaccttg
      961 tcatcttcag tatcatactg ttcgtataca tacttactga cattcatagg tatacatata
     1021 tacacatgta tatatatcgt atgctgcagc tttaaataat cggtgtcact acataagaac
     1081 acctttggtg gagggaacat cgttggtacc attgggcgag gtggcttctc ttatggcaac
     1141 cgcaagagcc ttgaacgcac tctcactacg gtgatgatca ttcttgcctc gcagacaatc
     1201 aacgtggagg gtaattctgc taggatctgc ccgccgggaa ggcgaacccg atcggatgca
     1261 tcctctctgc tgccatgatg ctgaagttgt cgttgaacat ggttgctgcc ggcgaggcgg
     1321 tcgagcaggc agtgcaggag gtgttggact cgggagtcag aacgggcgac ctgctcggct
     1381 cgagctccac ttcggaggtt ggcgacgcca ttgcgcttgc agttaaggaa gccttgcgca
     1441 ggcaatccgc agctggtctg agctagcctc gaggaccctt ctctttagac tattctactc
     1501 ttatgcacgt aaaaaattct aggaaatatg tattaactag gagtaaaata accggctagt
     1561 ggcattcata tagccgtctg tttacatcta catcacacat ttcgagtgta tatctcgcaa
     1621 cgttggcgtt aaataggcag tcaatggccc gaccattcta tggtgtttag gtcgatgcca
     1681 tctttgtaca gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga
     1741 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
     1801 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
     1861 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
     1921 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     1981 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     2041 ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg tttcgaggcc
     2101 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2161 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     2221 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2281 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2341 tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat tagaagaata
     2401 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2461 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2521 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2581 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
     2641 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     2701 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     2761 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     2821 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     2881 agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat
     2941 attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac
     3001 atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg
     3061 tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc agtgtcgact
     3121 agggttgctg ccatcggcct cgctcgcgtc tttgccggat agcaagagcg cctttggcct
     3181 ctacgagccc tgccacggct ctgcgcccga tctgcccgcc gggaaggcga acccgatcgg
     3241 atgcatcctc tctgctgcca tgatgctgaa gttgtcgttg aacatggttg ctgccggcga
     3301 ggcggtcgag caggcagtgc aggaggtgtt ggactcggga gtcagaacgg gcgacctgct
     3361 cggctcgagc tccacttcgg aggttggcga cgccattgcg cttgcagtta aggaagcctt
     3421 gcgcaggcaa tccgcagctg gtctgagcta gcctcgagga cccttctctt tagactattc
     3481 tactcttatg cacgtaaaaa attctaggaa atatgtatta actaggagta aaataaccgg
     3541 ctagtggcat tcatatagcc gtctgtttac atctacatca cacatttcga gtgtatatct
     3601 cgcaacgttg gcgttaaata ggcagtcaat ggcccgacca ttctatggtg tttaggtcga
     3661 tgccatcttt gtacgtttaa acgagctcga attctagcct ctgcaaagct ttcaagaaaa
     3721 tgcgggatca tctcgcaaga gagatctcct actttctccc tttgcaaacc aagttcgaca
     3781 actgcgtacg gcctgttcga aagatctacc accgctctgg aaagtgcctc atccaaaggc
     3841 gcaaatcctg atccaaacct ttttactcca cgcacggccc ctagggcctc tttaaaagct
     3901 tgaccgagag caatcccgca gtcttcagtg gtgtgatggt cgtctatgtg taagtcacca
     3961 atgcactcaa cgattagcga ccagccggaa tgcttggcca gagcatgtat catatggtcc
     4021 agaaacccta tacctgtgtg gacgttaatc acttgcgatt gtgtggcctg ttctgctact
     4081 gcttctgcct ctttttctgg gaagatcgag tgctctatcg ctaggggacc accctttaaa
     4141 gagatcgcaa tctgaatctt ggtttcattt gtaatacgct ttactagggc tttctgctct
     4201 gtcatctttg ccttcgttta tcttgcctgc tcatttttta gtatattctt cgaagaaatc
     4261 acattacttt atataatgta taattcatta tgtgataatg ccaatcgcta agaaaaaaaa
     4321 agagtcatcc gctaggtgga aaaaaaaaaa tgaaaatcat taccgaggca taaaaaaata
     4381 tagagtgtac tagaggaggc caagagtaat agaaaaagaa aattgcggga aaggactgtg
     4441 ttatgacttc cctgactaat gccgtgttca aacgatacct ggcagtgact cctagcgctc
     4501 accaagctct taaaacgaga attaagaaaa agtcgtcatc tttcgataag tttttcccac
     4561 agcaaagcaa tagtagaaaa aaacaatggg aaacgttgaa tgaagacaaa gcgtcgtggt
     4621 ttaaaaggaa atacgctcac gtacatgcta gggaacagga ccgtgcagcg gatccccggg
     4681 taccgagctc gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4741 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4801 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4861 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4921 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4981 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5041 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5101 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5161 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5221 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5281 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     5341 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5401 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5461 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5521 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     5581 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5641 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5761 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5821 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5881 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5941 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6001 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6061 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6121 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6181 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6241 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6301 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6361 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6421 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6481 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6541 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6601 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6661 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6721 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6781 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6841 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6901 ataggcgtat cacgaggccc tttcgtc
//
