LOCUS       sequence-79028-e        6063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid for integration of target sequence at HO locus, Kan 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6063)
  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 6063)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2001-06-15"; volume: "29(12)"; pages: "E59-9"
COMMENT     Sequence Label: HO-Poly-KanMX4-HO
FEATURES             Location/Qualifiers
     source          1..6063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            1070..2426
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1070..1413
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1414..2223
                     /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(1481..1500)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2229..2426
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     2996..3012
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2996..3012
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3026..3045)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3027..3045)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3052..3069)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3052..3068)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3061..3083)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3368..3387)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3500..3521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3675..3694)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3794..3816
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3854..3872)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3940..4044
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4045..4113,4114..4905)
                     /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(4263..4282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5076..5664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5565..5584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5818..5835
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5952..5973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    6028..6044
                     /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     6033..6055
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6052..6063,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6052..6063,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccact agtccgaggc ctcgagatcc gatatcgccg tggcggccgc
       61 cagctgaagc ttaattatcc tgggcacgag tgaaacaaag ctaaaacctt tatttagcat
      121 ggccattgaa tgtaacaatt atatatatcg caagcacaaa aaatcaagga gagagaacta
      181 ccactttgtt catgtgtaca atgttcatta tctccataag caaaaaaaaa aaaatagaaa
      241 acatatgcta taaggttgat attctcacga gtaagcggca cttgctactt attgacattg
      301 cagatttttg gctacagaaa tagtatatta gagattataa ttgctaatca aatcaaaata
      361 taaaattagt aaaccaaacc atttataccc ttccttagta gttatggatt gttttttaat
      421 gatatttctg caaaccaaag aaagattgtt atccagatag aatttagttt tgatattcat
      481 ttttttgttg aagattgaac gccatatctg ggcctcataa ttcaaaagac ggtgccatta
      541 tcggtagcgt ttcgcattgt actggatttc agaaatttca cagttgatga atcgaaaaga
      601 atggtctcat tgcaacacgt aaggttaaga tgtccctttt taccattata ggcaataaat
      661 gaatcataaa acgaccgtat actggtgaaa tagtagggag aacgagtacc tgtagtaaaa
      721 agtataaatc atagttaatc gggcaatgtc cctcgatcaa ggagtattgt gtcatgttcg
      781 agacaaacgc caacattttt gtttcttttg gacaaatgtt gtttgcattt atgatccgtt
      841 atattttgat ctaatgtaga gttgcacgta gttcttactg gcaaagaaat cgatgcatac
      901 caaaaaagaa taaaggtgat atttgatctt taccgtttag ttccaacgta aaattgtgcc
      961 tttggactta aaatggcgtc gtacgctgca ggtcgacgga tccccgggtt aattaaggcg
     1021 cgccagatct gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc
     1081 ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg
     1141 cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg
     1201 ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc
     1261 tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg
     1321 ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata
     1381 cagttctcac atcacatccg aacataaaca accatgggta aggaaaagac tcacgtttcg
     1441 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     1501 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     1561 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     1621 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     1681 gatgatgcat ggttactcac cactgcgatc cccggcaaaa cagcattcca ggtattagaa
     1741 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     1801 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     1861 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     1921 ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat
     1981 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2041 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2101 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2161 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2221 taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt
     2281 tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc
     2341 tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc
     2401 gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagtgt
     2461 cgaaaacgag ctcgaattcc tgggggaaca acttcacaga atgttttgtc atattgtcga
     2521 agtggtcaca aaacaagaga agttccgcca attataaaaa gggaacccgt atatttcagc
     2581 ttcacggatg atttccaggg tgagagtact gtatatgggc ttacgataga aggccataaa
     2641 aatttcttgc ttggcaacaa aatagaagtg aaatcatgtc gaggctgctg tgtgggagaa
     2701 cagcataaaa tatcacaaaa aaagaatcta aaacactgtg ttgcttgtcc cagaaaggga
     2761 atcaagtatt tttataaaga ttggagtggt aaaaatcgag tatgtgctag atgctatgga
     2821 agatacaaat tcagcggtca tcactgtata aattgcaagt atgtaccaga agcacgtgaa
     2881 gtgaaaaagg caaaagacaa aggcgaaaaa ttgggcatta cgcccgaagg tttgccagtt
     2941 aaaggaccag agtgtataaa atgtggcgga atcttacagt ggcctatgcg gccgctctag
     3001 aactagtgga tcgatcccca attcgcccta tagtgagtcg tattacaatt cactggccgt
     3061 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     3121 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3181 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     3241 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     3301 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3361 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3421 caagctctaa atcggggtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3481 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3541 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3601 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     3661 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     3721 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3781 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3841 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3901 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3961 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4021 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4081 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4141 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4201 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4261 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4321 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4381 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4441 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4501 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4561 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4621 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4681 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4741 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4801 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4861 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4921 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4981 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5041 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5221 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5461 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5641 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5701 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5761 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5821 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     5881 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     5941 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     6001 cttccgcggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     6061 gct
//
