LOCUS       sequence-255871-        5349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for ScCUP1 promoter regulated N-terminal fused Nui-HA prey 
            expression, centromeric ARS plasmid, 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 5349)
  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 5349)
  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: pCUP1pNuiHA kanMX CEN
FEATURES             Location/Qualifiers
     source          1..5349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        191..471
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             complement(706..1515)
                     /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     1429..1448
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1516..1859)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        2076..2310
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             2482..2508
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2531..2547)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2532..2548)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2576..2594)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2615..2631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2615..2631)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2628..2650)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2639..2655
                     /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)."
     promoter        complement(join(2663..2669,2670..2687,2688..2693))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2708..2729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2846..2863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3017..3605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3097..3116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3776..4567,4568..4636))
                     /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     4399..4418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4637..4741)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4778..5281
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5322..5340
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatagatc acgacattac tatatatata atataggaag catttaatag aacagcatcg
      241 taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat
      301 tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga
      361 ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga
      421 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt
      481 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
      541 gccgcagaat gtgcgcactt aacttcgcat ctgggcagat gatgtcgagg cgaaaaaaaa
      601 tataaatcac gctaacattt gattaaaata gaacaactac aatataaaaa aactatacaa
      661 atgacaagtt cttgaaaaca agaatctttt tattgtcagt actgattaga aaaactcatc
      721 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
      781 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
      841 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
      901 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
      961 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     1021 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     1081 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     1141 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     1201 gaatgctgtt ttgccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     1261 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     1321 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     1381 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     1441 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg aaacgtgagt
     1501 cttttcctta cccatggttg tttatgttcg gatgtgatgt gagaactgta tcctagcaag
     1561 attttaaaag gaagtatatg aaagaagaac ctcagtggca aatcctaacc ttttatattt
     1621 ctctacaggg gcgcggcgtg gggacaattc aacgcgtctg tgaggggagc gtttccctgc
     1681 tcgcaggtct gcagcgagga gccgtaattt ttgcttcgcg ccgtgcggcc atcaaaatgt
     1741 atggatgcaa atgattatac atggggatgt atgggctaaa tgtacgggcg acagtcacat
     1801 catgcccctg agctgcgcac gtcaagactg tcaaggaggg tattctgggc ctccatgtcg
     1861 ctggccgggt gacccggcgg ggacgaggca agctaaacag atctggacga tcccattacc
     1921 gacatttggg cgctatacgt gcatatgttc atgtatgtat ctgtatttaa aacacttttg
     1981 tattattttt cctcatatat gtgtataggt ttatacggat gatttaatta ttacttcacc
     2041 accctttatt tcaggctgat atcttagcct tgttactagt tagaaaaaga catttttgct
     2101 gtcagtcact gtcaagagat tcttttgctg gcatttcttc tagaagcaaa aagagcgatg
     2161 cgtcttttcc gctgaaccgt tccagcaaaa aagactacca acgcaatatg gattgtcaga
     2221 atcatataaa agagaagcaa ataactcctt gtcttgtatc aattgcatta taatatcttc
     2281 ttgttagtgc aatatcatat agaagtcatc gaaatagata ttaagaaaaa caaactgtaa
     2341 cgaatttcga ccggcggtat gcagattttc gtcaagactt tgaccggtaa aaccataaca
     2401 ttggaagttg aatcttccga taccatcgac aacgttaagt cgaaaattca agacaaggaa
     2461 gggatccctg ggtctggggc ttacccatac gatgttcctg actatgcggg gaattcgata
     2521 tcaagcttat cgataccgtc gacctcgagg gggggcccgg taccagcttt tgttcccttt
     2581 agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2641 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2701 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2761 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2821 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2881 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2941 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3001 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3061 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3121 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3181 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3241 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3301 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3361 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3421 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3481 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3541 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3601 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3661 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3721 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3781 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3841 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3901 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3961 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4021 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4081 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4141 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4201 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4261 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4321 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4381 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4441 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4501 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4561 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4621 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4681 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4741 gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc acgtgctata aaaataatta
     4801 taatttaaat tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta
     4861 aagaaaaaat agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa
     4921 tactaccttt tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg
     4981 tctgtgtaga agaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg
     5041 aatgtatatc tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt
     5101 ttgttgaaat tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc
     5161 ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa
     5221 attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat
     5281 ccgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5341 cctttcgtc
//
