LOCUS       sequence-255835-        4520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR template for genomic ScCUP1 promoter regulated N-terminal Nui-HA
            prey fusion, kanMX marker confers resistance to geneticin in S. 
            cerevisiae .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4520)
  AUTHORS   Dunkler A, Muller J, Johnsson N
  TITLE     Detecting protein-protein interactions with the Split-Ubiquitin 
            sensor.
  JOURNAL   Methods Mol Biol. 2012;786:115-30. doi: 10.1007/978-1-61779-292-2_7.
  PUBMED    21938623
REFERENCE   2  (bases 1 to 4520)
  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: "Methods Mol
            Biol."; date: "2012"; volume: "786:115-30. doi"; pages: " 
            10.1007/978-1-61779-292-2_7"
COMMENT     Sequence Label: pFA CUP1p-NuiHA kanMX
FEATURES             Location/Qualifiers
     source          1..4520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..42
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     24..41
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     gene            complement(115..1471)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(115..312)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(318..1127)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RY*DLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1041..1060
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1128..1471)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        1688..1922
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             2094..2120
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2197..2216)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2198..2216)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2303..2320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2474..3062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2554..2573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3233..4024,4025..4093))
                     /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     3856..3875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4094..4198)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4266..4284
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4322..4344)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4444..4463
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 accttatgta tcatacacat acgatttagg tgacactata gaacgcggcc gccagctgaa
       61 gcttcgtacg ctgcaggtcg acactggatg gcggcgttag tatcgaatcg acagcagtat
      121 agcgaccagc attcacatac gattgacgca tgatattact ttctgcgcac ttaacttcgc
      181 atctgggcag atgatgtcga ggcgaaaaaa aatataaatc acgctaacat ttgattaaaa
      241 tagaacaact acaatataaa aaaactatac aaatgacaag ttcttgaaaa caagaatctt
      301 tttattgtca gtactgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
      361 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
      421 tcaccgaggc agttccatag gatggcaaga tcctagtatc ggtctgcgat tccgactcgt
      481 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
      541 tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag
      601 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
      661 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
      721 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
      781 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttgccggg gatcgcagtg
      841 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
      901 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
      961 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     1021 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     1081 ttggaattta atcgcggcct cgaaacgtga gtcttttcct tacccatggt tgtttatgtt
     1141 cggatgtgat gtgagaactg tatcctagca agattttaaa aggaagtata tgaaagaaga
     1201 acctcagtgg caaatcctaa ccttttatat ttctctacag gggcgcggcg tggggacaat
     1261 tcaacgcgtc tgtgagggga gcgtttccct gctcgcaggt ctgcagcgag gagccgtaat
     1321 ttttgcttcg cgccgtgcgg ccatcaaaat gtatggatgc aaatgattat acatggggat
     1381 gtatgggcta aatgtacggg cgacagtcac atcatgcccc tgagctgcgc acgtcaagac
     1441 tgtcaaggag ggtattctgg gcctccatgt cgctggccgg gtgacccggc ggggacgagg
     1501 caagctaaac agatctggac gatcccatta ccgacatttg ggcgctatac gtgcatatgt
     1561 tcatgtatgt atctgtattt aaaacacttt tgtattattt ttcctcatat atgtgtatag
     1621 gtttatacgg atgatttaat tattacttca ccacccttta tttcaggctg atatcttagc
     1681 cttgttacta gttagaaaaa gacatttttg ctgtcagtca ctgtcaagag attcttttgc
     1741 tggcatttct tctagaagca aaaagagcga tgcgtctttt ccgctgaacc gttccagcaa
     1801 aaaagactac caacgcaata tggattgtca gaatcatata aaagagaagc aaataactcc
     1861 ttgtcttgta tcaattgcat tataatatct tcttgttagt gcaatatcat atagaagtca
     1921 tcgaaataga tattaagaaa aacaaactgt aacgaatttc gaccggcggt atgcagattt
     1981 tcgtcaagac tttgaccggt aaaaccataa cattggaagt tgaatcttcc gataccatcg
     2041 acaacgttaa gtcgaaaatt caagacaagg aagggatccc tgggtctggg gcttacccat
     2101 acgatgttcc tgactatgcg ggcgcgccga cgacagagaa ttcatcgatg atatcagatc
     2161 cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt
     2221 cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2281 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2341 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2401 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2461 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2521 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2581 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2641 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2701 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2761 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2821 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2881 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2941 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3001 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3061 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3121 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3181 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3241 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3301 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3361 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3421 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3481 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3541 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3601 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3661 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3721 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3781 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3841 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3901 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3961 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4021 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4081 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4141 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4201 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4261 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     4321 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4381 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     4441 atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac
     4501 gcggctacaa ttaatacata
//
