LOCUS       sequence-79029-e        8488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid for integration of target sequence at HO locus, URA3 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8488)
  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 8488)
  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-hisG-URA3-hisG-poly-HO
FEATURES             Location/Qualifiers
     source          1..8488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2195..2415
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2416..3219
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     5421..5437
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5421..5437
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5451..5470)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5452..5470)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5477..5494)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5477..5493)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5486..5508)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5793..5812)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5925..5946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6100..6119)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6219..6241
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6279..6297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6365..6469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6470..6538,6539..7330)
                     /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(6688..6707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7501..8089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7990..8009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8243..8260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8377..8398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    8453..8469
                     /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     8458..8480
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8477..8488,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8477..8488,1..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 tcttccagtg gtgcatgaac
     1021 gcatgagaaa gcccccggaa gatcatcttc cgggggcttt ttttttggcg cgcgatacag
     1081 accggttcag acaggataaa gaggaacgca gaatgttaga caacacccgc ttacgcatag
     1141 ctattcagaa atcaggccgt ttaagcgatg attcacgaga attgctggcc cgctgcggca
     1201 taaaaattaa tttacacact cagcgcctga ttgcgatggc ggaaaacatg ccgattgata
     1261 tcctgcgcgt gcgtgatgat gacattccgg gtctggtaat ggatggcgtg gtcgatctcg
     1321 gtattatcgg cgaaaacgtg ctggaagaag agctactcaa ccgccgcgca cagggcgaag
     1381 atccacgcta tttaaccctg cgccgtcttg acttcggcgg ctgccgttta tcgctggcaa
     1441 caccggttga cgaagcctgg gacggcccgg ccgcgctgga cggtaaacgt atcgctacct
     1501 catatccgca cctcctcaaa cgctacctcg accagaaagg cgtctctttt aaatcgtgtc
     1561 tgttaaatgg ttctgtcgaa gtcgcgccgc gcgcggggct ggccgacgct atctgcgatt
     1621 tggtctctac cggcgcgacg cttgaagcta acggcctgcg tgaagtcgaa gttatctacc
     1681 gctctaaagc ctgtctgatt cagcgcgacg gtgagatggc acagagcaag caagagctga
     1741 tcgataaatt gctgacccgt attcagggcg tgattcaggc gcgcgaatcg aaatacatca
     1801 tgatgcacgc gccaagtgaa cgcctggaag aggttatcgc cctgctgcca ggcgccgaaa
     1861 ggccgacaat tctgccgctg gcaggcgagc aacagcgcgt ggcgatgcac atggtcagca
     1921 gcgaaacgtt gttctgggaa accatggaga aactgaaagc gcttggcgcc agctcgattc
     1981 tggtactgcc gatcgagaag atgatggagt gatctgacgc ctgatggcgc tgcgcttatc
     2041 aggcctacgt aatgcgttga tattttgggt tctgtaggcc ggataaggcg gaaccctgtg
     2101 atggagtaaa gaccatgagc ttcaataccc tgattgactg gaacagcgga tctgaattaa
     2161 ttctcatgtt tgacagctta tcatcgataa gcttttcaat tcaattcatc attttttttt
     2221 tattcttttt tttgatttcg gtttccttga aatttttttg attcggtaat ctccgaacag
     2281 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     2341 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacctgcagg
     2401 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     2461 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     2521 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     2581 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     2641 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct
     2701 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     2761 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     2821 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     2881 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     2941 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     3001 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagacgc attgggtcaa
     3061 cagtatagaa ccgtggatga tgtggtctct acaggatctg acattattat tgttggaaga
     3121 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     3181 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     3241 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccggg
     3301 aatctcggtc gtaatgattt ttataatgac gaaaaaaaaa aaattggaaa gaaaaagctt
     3361 taatgcggta gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct
     3421 aacaatgcgc tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg
     3481 gttatgccgg tactgccggg cctcttgcgg gatatcgtcc attccgacag catcgccagt
     3541 cactatggcg tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc acccgttctc
     3601 ggagcactgt ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct acttggagcc
     3661 actatcgact acgcgatcat ggcgaccaca cccgtcctgt ggatcttcca gtggtgcatg
     3721 aacgcatgag aaagcccccg gaagatcatc ttccgggggc tttttttttg gcgcgcgata
     3781 cagaccggtt cagacaggat aaagaggaac gcagaatgtt agacaacacc cgcttacgca
     3841 tagctattca gaaatcaggc cgtttaagcg atgattcacg agaattgctg gcccgctgcg
     3901 gcataaaaat taatttacac actcagcgcc tgattgcgat ggcggaaaac atgccgattg
     3961 atatcctgcg cgtgcgtgat gatgacattc cgggtctggt aatggatggc gtggtcgatc
     4021 tcggtattat cggcgaaaac gtgctggaag aagagctact caaccgccgc gcacagggcg
     4081 aagatccacg ctatttaacc ctgcgccgtc ttgacttcgg cggctgccgt ttatcgctgg
     4141 caacaccggt tgacgaagcc tgggacggcc cggccgcgct ggacggtaaa cgtatcgcta
     4201 cctcatatcc gcacctcctc aaacgctacc tcgaccagaa aggcgtctct tttaaatcgt
     4261 gtctgttaaa tggttctgtc gaagtcgcgc cgcgcgcggg gctggccgac gctatctgcg
     4321 atttggtctc taccggcgcg acgcttgaag ctaacggcct gcgtgaagtc gaagttatct
     4381 accgctctaa agcctgtctg attcagcgcg acggtgagat ggcacagagc aagcaagagc
     4441 tgatcgataa attgctgacc cgtattcagg gcgtgattca ggcgcgcgaa tcgaaataca
     4501 tcatgatgca cgcgccaagt gaacgcctgg aagaggttat cgccctgctg ccaggcgccg
     4561 aaaggccgac aattctgccg ctggcaggcg agcaacagcg cgtggcgatg cacatggtca
     4621 gcagcgaaac gttgttctgg gaaaccatgg agaaactgaa agcgcttggc gccagctcga
     4681 ttctggtact gccgatcgag aagatgatgg agtgatctga cgcctgatgg cgctgcgctt
     4741 atcaggccta cgtaatgcgt tgatattttg ggttctgtag gccggataag gcggaaccct
     4801 gtgatggagt aaagaccatg agcttcaata ccctgattga ctggaacagc ggatccccgg
     4861 ggttaattaa ggcgcgccag atctgtttaa acgagctcga attcctgggg gaacaacttc
     4921 acagaatgtt ttgtcatatt gtcgaagtgg tcacaaaaca agagaagttc cgccaattat
     4981 aaaaagggaa cccgtatatt tcagcttcac ggatgatttc cagggtgaga gtactgtata
     5041 tgggcttacg atagaaggcc ataaaaattt cttgcttggc aacaaaatag aagtgaaatc
     5101 atgtcgaggc tgctgtgtgg gagaacagct taaaatatca caaaaaaaga atctaaaaca
     5161 ctgtgttgct tgtcccagaa agggaatcaa gtatttttat aaagattgga gtggtaaaaa
     5221 tcgagtatgt gctagatgct atggaagata caaattcagc ggtcatcact gtataaattg
     5281 caagtatgta ccagaagcac gtgaagtgaa aaaggcaaaa gacaaaggcg aaaaattggg
     5341 cattacgccc gaaggtttgc cagttaaagg accagagtgt ataaaatgtg gcggaatctt
     5401 acagtggcct atgcggccgc tctagaacta gtggatcgat ccccaattcg ccctatagtg
     5461 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     5521 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     5581 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     5641 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     5701 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5761 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5821 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggtgggccat cgccctgata
     5881 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5941 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     6001 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     6061 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     6121 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6181 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6241 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     6301 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     6361 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6421 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6481 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6541 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6601 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6661 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6721 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6781 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6841 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6901 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6961 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7021 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7081 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7141 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7201 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7261 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7321 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7381 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7441 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7501 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7561 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7621 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     7681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7861 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     7921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7981 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8041 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8101 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     8161 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     8221 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     8281 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     8341 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     8401 ggcaccccag gctttacact ttatgcttcc gcggctcgta tgttgtgtgg aattgtgagc
     8461 ggataacaat ttcacacagg aaacagct
//
