LOCUS       sequence-487259-        8002 bp    DNA     circular SYN 26-SEP-2025
DEFINITION  High-copy episomal vector for galactose-inducible expression of 
            TaHKT1 protein in Saccharomyces cerevisiae..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8002)
  AUTHORS   Schachtman DP, Schroeder JI
  TITLE     Structure and transport mechanism of a high-affinity potassium 
            uptake transporter from higher plants.
  JOURNAL   Nature. 1994 Aug 25;370(6491):655-8. doi: 10.1038/370655a0.
  PUBMED    8065452
REFERENCE   2  (bases 1 to 8002)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nature"; date: "1994-08-25- 25"; volume: "370"; issue: "6491"; 
            pages: "655-8"
COMMENT     Sequence Label: TaHKT1_pYES2
FEATURES             Location/Qualifiers
     source          1..8002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(837..1084)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     1050..1068
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     3309..3325
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3310..3326
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3341..3360)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3342..3360)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3392..3833)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(3398..3421)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    3716..3833
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     rep_origin      3855..4368
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3942..3961)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4152..4173
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4379..5256
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5857..6072
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             6073..6876
                     /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"
     CDS             join(6972..7040,7041..7832)
                     /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(7190..7209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagggccg cagcttgcaa
      841 attaaagcct tcgagcgtcc caaaaccttc tcaagcaagg ttttcagtat aatgttacat
      901 gcgtacacgc gtctgtacag aaaaaaaaga aaaatttgaa atataaataa cgttcttaat
      961 actaacataa ctataaaaaa ataaataggg acctagactt caggttgtct aactccttcc
     1021 ttttcggtta gagcggatgt ggggggaggg cgtgaatgta agcgtgacat aactaattac
     1081 atgatgcggc cgctcgagct acgtcagggt tttttttttt tttttttttt tttttttttt
     1141 tttttttttt tttttttttt tttttttttt gataatcata gacttttcta tgaaaccggg
     1201 taaaaataca ttatctgcag atatacatgc tctataaatg aagtcgagtt ttcaaactca
     1261 atttgatgaa aaatacaaat gatcaccatc ccaaatgaca tctaaatcat caatctcagc
     1321 gttgtccgca ttggtgatct ttgtgaagtt gcagcctcca aacttctcac ttgcaagtct
     1381 tagaagctct tctatggatg atggtaagag tatcaacttc ccgttcttgc tctcagacat
     1441 catgtggatg gtaactctct tttcttctcg tttgcatggc ttgatgattt ggttgcttga
     1501 gctgcaatat ggatcttttt tcgatgaatc gttagcgtaa cttcttccag cggtttcact
     1561 acttgatctt tccactttgg ctctctctat cttttgttcc ttaagtatct tcttaaccat
     1621 ttcagtatct tccttgtgaa tcgcatccat tagagcagtg tcggaagtcg gattcgtaac
     1681 atctaaacca tagccatctt ttcttgaatc tctccactct tcccatccca tgcttttgaa
     1741 atctctgttt tcgtgaccac tagtattcga atcatctatt gctattgact gttgccctct
     1801 aagtttcatg aagagattgt tcatgatgac tcgtccatcg tcagcatgag catgcatcgc
     1861 actcatcagc gatgttctgc ttatccgcag tatttgagat agctcggttg ttcttactgt
     1921 gaatggttgt ggtctatagt ataaaactcc aacctctcca aatgtttctc caattactgc
     1981 tttcccttga aactgatcat gtccatcaac gtagacagtg aagtccactg ctcctgacac
     2041 cagaatgtaa agatcagtag gagcttcgtt ttgtagaatt atatcttctt ttggtgggaa
     2101 atactcagcg tctatatctg aaaccaattg aaagaggaag ttacgagaaa ctccttgaaa
     2161 gaggtaaatg ttgtgaacaa tggggaagaa taaatagttt gcaatgcttg accggattgc
     2221 ttttggcaga ttgttcaagg tctcttgttg tttcaagccc tctgttttga actttaagca
     2281 aatgtgtgat aacatttgat cttgtatgtc atgtgggagt tgatttctag aagcaaactc
     2341 tgaagcagct ctcactgaat ccctaaaggt tctggttcgg ctagtccaat gaacgacgag
     2401 gttggtcata tttccaatga ggtaagctgt caaaccgagg ttgaacatca tgaagaaaat
     2461 gtcaaaaagc atttctcttg ggttctcagc atgaaaatct ccatatcccg tggtcgttaa
     2521 tgtcgtaatg gaccagtaaa gagcagtcac atatctattc catagacttg cttctttgaa
     2581 atttggatac acagctccaa tccatgtctt tcttggatta ggatatctat ctgcaatcag
     2641 gtagttgaaa catccagcac aatgtatagc gaacaaagtg accgaaatga gttttgtgca
     2701 acgtatccag aaatagttga aacggatatc tttctcaagc cttgcaaata gcgagctaac
     2761 tcgccggaga cgccataacc tgagcatgct aagaattctg aatcctagtt cgcttccgtt
     2821 gtagttaaac aagaggctta gtggctgaaa tggtgctgtg gaacaaacat cgaaagcgaa
     2881 ccacgtcgaa aggtacctta ttgctatttt cttaggactg tcaactagaa gataggagtg
     2941 gctatcgaga taagcgacga agaaggtgag aataatatca atggcgaaga agccattaac
     3001 aatgttgtcg atgatgaaaa tcgcgtcttt tttataggtg atgaaagcaa attgaaatgg
     3061 gcaaatccaa gctgagtaaa taactagtaa tactagccac atctcccacg ctctgtaacg
     3121 tggattaaaa ggagagatta tgtgtttgcg gagcttagta gattggttga tcctggctcc
     3181 aagagatggt agaagatcgg cagagaggaa actactctgt tttatggtgt ctatattgta
     3241 ttcctcgacg cagaatcttt cgaagaaatt tcgagtccaa gagatcgaca tcttttccct
     3301 gacgtagctc gaggtcgacg gtatcgataa gcttaatatt ccctatagtg agtcgtatta
     3361 cagctgctag tagtccgatc cggggttttt tctccttgac gttaaagtat agaggtatat
     3421 taacaatttt ttgttgatac ttttattaca tttgaataag aagtaataca aaccgaaaat
     3481 gttgaaagta ttagttaaag tggttatgca gtttttgcat ttatatatct gttaatagat
     3541 caaaaatcat cgcttcgctg attaattacc ccagaaataa ggctaaaaaa ctaatcgcat
     3601 tatcatccta tggttgttaa tttgattcgt tcatttgaag gtttgtgggg ccaggttact
     3661 gccaattttt cctcttcata accataaaag ctagtattgt agaatcttta ttgttcggag
     3721 cagtgcggcg cgaggcacat ctgcgtttca ggaacgcgac cggtgaagac gaggacgcac
     3781 ggaggagagt cttccttcgg agggctgtca cccgctcggc ggcttctaat ccgtggacta
     3841 gtggatcgat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3901 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3961 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4021 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4081 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4141 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4201 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4261 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aaatatttgc
     4321 ttatacaatc ttcctgtttt tggggctttt ctgattatca accggggtgg agcttcccat
     4381 tgcgaatacc gcttccacaa acattgctca aaagtatctc tttgctatat atctctgtgc
     4441 tatatcccta tataacctac ccatccacct ttcgctcctt gaacttgcat ctaaactcga
     4501 cctctacatt ttttatgttt atctctagta ttactcttta gacaaaaaaa ttgtagtaag
     4561 aactattcat agagtgaatc gaaaacaata cgaaaatgta aacatttcct atacgtagta
     4621 tatagagaca aaatagaaga aaccgttcat aattttctga ccaatgaaga atcatcaacg
     4681 ctatcacttt ctgttcacaa agtatgcgca atccacatcg gtatagaata taatcgggga
     4741 tgcctttatc ttgaaaaaat gcacccgcag cttcgctagt aatcagtaaa cgcgggaagt
     4801 ggagtcaggc tttttttatg gaagagaaaa tagacaccaa agtagccttc ttctaacctt
     4861 aacggaccta cagtgcaaaa agttatcaag agactgcatt atagagcgca caaaggagaa
     4921 aaaaagtaat ctaagatgct ttgttagaaa aatagcgctc tcgggatgca tttttgtaga
     4981 acaaaaaaga agtatagatt ctttgttggt aaaatagcgc tctcgcgttg catttctgtt
     5041 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     5101 gttttacaaa aatgaagcac agattcttcg ttggtaaaat agcgctttcg cgttgcattt
     5161 ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca
     5221 tttttgttct acaaaatgaa gcacagatgc ttcgttaaca aagatatgct attgaagtgc
     5281 aagatggaaa cgcagaaaat gaaccgggga tgcgacgtgc aagattacct atgcaataga
     5341 tgcaatagtt tctccaggaa ccgaaataca tacattgtct tccgtaaagc gctagactat
     5401 atattattat acaggttcaa atatactatc tgtttcaggg aaaactccca ggttcggatg
     5461 ttcaaaattc aatgatgggt aacaagtacg atcgtaaatc tgtaaaacag tttgtcggat
     5521 attaggctgt atctcctcaa agcgtattcg aatatcattg agaagctgca gcgtcacatc
     5581 ggataataat gatggcagcc attgtagaag tgccttttgc atttctagtc tctttctcgg
     5641 tctagctagt tttactacat cgcgaagata gaatcttaga tcacactgcc tttgctgagc
     5701 tggatcaata gagtaacaaa agagtggtaa ggcctcgtta aaggacaagg acctgagcgg
     5761 aagtgtatcg tacagtagac ggagtatact aggtatagtc tatagtccgt ggaattaatt
     5821 ctcatgtttg acagcttatc atcgataagc tagcttttca attcatcatt ttttttttat
     5881 tctttttttt gatttcggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
     5941 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
     6001 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac ctgcaggaaa
     6061 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     6121 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     6181 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     6241 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     6301 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac
     6361 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     6421 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     6481 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     6541 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     6601 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     6661 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagacgcatt gggtcaacag
     6721 tatagaaccg tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga
     6781 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     6841 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     6901 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccattgaa
     6961 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7021 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7081 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7141 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     7201 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     7261 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     7321 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7381 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7441 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7501 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7561 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7621 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7681 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7741 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7801 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7861 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7921 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7981 tagaaaagat caaaggatct tc
//
