LOCUS       Exported                6760 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  yeast plasmid for integration of target sequence at HO locus, Kan 
            selection, pMET inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    HO-pMET-poly-KanMX4-HO
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6760)
  AUTHORS   Voth WP, Richards JD, Shaw JM, Stillman DJ
  TITLE     Yeast vectors for integration at the HO locus.
  JOURNAL   Nucleic Acids Res. 2001 Jun 15;29(12):E59-9.
  PUBMED    11410682
REFERENCE   2  (bases 1 to 6760)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6760
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     206..224
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     262..278
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     263..279
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(313..329)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(313..329)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(330..679)
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     gene            779..2135
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        779..1122
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1123..1932
                     /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(1190..1209)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1938..2135
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2743..2762)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2744..2762)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2769..2786)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2769..2785)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2778..2800)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3085..3104)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3217..3238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3392..3411)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3511..3533
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3571..3589)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3657..3761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3762..4622
                     /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     complement(3980..3999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4793..5381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5282..5301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5535..5552
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5669..5690
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    5745..5761
                     /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     5750..5772
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5769..5785
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5769..5785
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gtacccgagc gtcccaaaac cttctcaagc aaggttttca gtataatgtt acatgcgtac
       61 acgcgtttgt acagaaaaaa aagaaaaatt tgaaatataa ataacgttct taatactaac
      121 ataactatta aaaaaaataa atagggacct agacttcagg ttgtctaact ccttcctttt
      181 cggttagagc ggatgtggga ggagggcgtg aatgtaagcg tgacataact aattacatga
      241 tatcgacaaa ggaaaagggg ctcgaggtcg acggtatcga taagcttgat atcgaattcc
      301 tgcagcccgg gggatccact agttctagag tatggatggg ggtaatagaa ttgtatctat
      361 gtatctgacg accctgtatt acacgaagga aatagtagat gaaaagacaa gagagcaaga
      421 aaaaggaaaa acttcctttc taacagacgc tttacttaac cttatatata tcttattttt
      481 ttcatcgagc gtgttcaatt ggacaaggtg ccaccttttc gacacttcgg ttattatgtt
      541 acacagtttt catgaggatg gcgccttgac taacttaatt agtcatttgc caaccaccac
      601 agttccccaa tatcgacagc ttcacgtgcc atttgccatt ttgcgatcat gtgacctcaa
      661 gagaaaaagc aaaaaataat gagagctaag ggattcgaac ccttgcatcc ggaggggtta
      721 attaaggcgc gccagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga
      781 catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg
      841 tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg catccataca
      901 ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca
      961 gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa
     1021 tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt
     1081 gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa ggaaaagact
     1141 cacgtttcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1201 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     1261 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1321 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1381 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggcaaaac agcattccag
     1441 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1501 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1561 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1621 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc
     1681 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     1741 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     1801 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     1861 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     1921 gagtttttct aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg
     1981 tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt
     2041 tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat
     2101 gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc
     2161 atccagtgtc gaaaacgagc tcgaattaat tcctggggga acaacttcac agaatgtttt
     2221 gtcatattgt cgaagtggtc acaaaacaag agaagttccg ccaattataa aaagggaacc
     2281 cgtatatttc agcttcacgg atgatttcca gggtgagagt actgtatatg ggcttacgat
     2341 agaaggccat aaaaatttct tgcttggcaa caaaatagaa gtgaaatcat gtcgaggctg
     2401 ctgtgtggga gaacagcata aaatatcaca aaaaaagaat ctaaaacact gtgttgcttg
     2461 tcccagaaag ggaatcaagt atttttataa agattggagt ggtaaaaatc gagtatgtgc
     2521 tagatgctat ggaagataca aattcagcgg tcatcactgt ataaattgca agtatgtacc
     2581 agaagcacgt gaagtgaaaa aggcaaaaga caaaggcgaa aaattgggca ttacgcccga
     2641 aggtttgcca gttaaaggac cagagtgtat aaaatgtggc ggaatcttac agtggcctat
     2701 gcggccgctc tagctagaac tagtggatcg atccccaatt cgccctatag tgagtcgtat
     2761 tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2821 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2881 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     2941 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3001 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3061 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3121 tcgccggctt tccccgtcaa gctctaaatc ggggtgggcc atcgccctga tagacggttt
     3181 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3241 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3301 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3361 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3421 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3481 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3541 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3601 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3661 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3721 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3781 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3841 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3901 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3961 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4021 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4081 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4141 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4201 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4261 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4321 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4381 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4441 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4501 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4561 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4621 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4681 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4741 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4801 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4861 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4921 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4981 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5041 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5101 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5161 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     5221 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5281 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5341 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5401 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5461 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5521 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     5581 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     5641 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     5701 aggctttaca ctttatgctt ccgcggctcg tatgttgtgt ggaattgtga gcggataaca
     5761 atttcacaca ggaaacagct atgaccatga ttacgccact agtccgaggc ctcgagatcc
     5821 gatatcgccg tggcggccgc cagctgaagc ttaattatcc tgggcacgag tgaaacaaag
     5881 ctaaaacctt tatttagcat ggccattgaa tgtaacaatt atatatatcg caagcacaaa
     5941 aaatcaagga gagagaacta ccactttgtt catgtgtaca atgttcatta tctccataag
     6001 caaaaaaaaa aaaatagaaa acatatgcta taaggttgat attctcacga gtaagcggca
     6061 cttgctactt attgacattg cagatttttg gctacagaaa tagtatatta gagattataa
     6121 ttgctaatca aatcaaaata taaaattagt aaaccaaacc atttataccc ttccttagta
     6181 gttatggatt gttttttaat gatatttctg caaaccaaag aaagattgtt atccagatag
     6241 aatttagttt tgatattcat ttttttgttg aagattgaac gccatatctg ggcctcataa
     6301 ttcaaaagac ggtgccatta tcggtagcgt ttcgcattgt actggatttc agaaatttca
     6361 cagttgatga atcgaaaaga atggtctcat tgcaacacgt aaggttaaga tgtccctttt
     6421 taccattata ggcaataaat gaatcataaa acgaccgtat actggtgaaa tagtagggag
     6481 aacgagtacc tgtagtaaaa agtataaatc atagttaatc gggcaatgtc cctcgatcaa
     6541 ggagtattgt gtcatgttcg agacaaacgc caacattttt gtttcttttg gacaaatgtt
     6601 gtttgcattt atgatccgtt atattttgat ctaatgtaga gttgcacgta gttcttactg
     6661 gcaaagaaat cgatgcatac caaaaaagaa taaaggtgat atttgatctt taccgtttag
     6721 ttccaacgta aaattgtgcc tttggactta aaatggcgtc
//
