LOCUS       sequence-432118-        6601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Component of yeast three hybrid system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6601)
  AUTHORS   Bernstein DS, Buter N, Stumpf C, Wickens M
  TITLE     Analyzing mRNA-protein complexes using a yeast three-hybrid system.
  JOURNAL   Methods. 2002 Feb;26(2):123-41. doi: 10.1016/S1046-2023(02)00015-4.
  PUBMED    12054889
REFERENCE   2  (bases 1 to 6601)
  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"; doi: "10"; journalName: 
            "Methods"; date: "2002-02"; volume: "26"; issue: "2"; pages: 
            "123-41"
COMMENT     Sequence Label: p3HR2
FEATURES             Location/Qualifiers
     source          1..6601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      69..1410
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    401..448
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        1437..1541
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1542..1610,1611..2402)
                     /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(1760..1779)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2573..3161
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3062..3081
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3315..3332
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(3485..3490,3491..3508,3509..3515)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3523..3539
                     /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     3528..3550
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3547..3563
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3547..3563
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3582..3602
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3584..3602
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3639..3655
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3657..4062
                     /gene="RPR1"
                     /label=RPR1 promoter
                     /note="promoter for the S. cerevisiae RNase P RNA gene"
     misc_RNA        4163..4181
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4202..4220
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      4253..4315
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
     primer_bind     complement(4606..4625)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4607..4625)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4635..4652)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4635..4651)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4644..4666)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4796..5251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4883..4902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5093..5114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5382..6185)
                     /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"
     promoter        complement(6186..6406)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6432..6451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6551..6573
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
      121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
      181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
      241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
      301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
      361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
      421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
      481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
      541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
      601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
      661 acatttttta tgtttatctc tagtattact ctttagaaaa aaaattgtag taagaactat
      721 tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga
      781 gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca
      841 ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt
      901 tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc
      961 aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga
     1021 cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag
     1081 taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa
     1141 aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa
     1201 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta
     1261 caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc
     1321 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg
     1381 ttctacaaaa tgaagcacag atgcttcgtt caggtggcac ttttcgggga aatgtgcgcg
     1441 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1501 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1561 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1621 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1681 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1741 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     1801 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     1861 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     1921 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1981 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2041 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2101 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2161 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2221 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2281 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2341 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2401 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2461 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2521 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2581 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2641 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2701 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2761 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2821 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2881 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2941 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3001 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3061 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3121 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3181 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3241 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3301 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     3361 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     3421 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta cctcactcat taggcacccc
     3481 aggctttaca ctttatgctt ccggctccta tgttgtgtgg aattgtgagc ggataacaat
     3541 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     3601 gggaacaaaa gctggagctc caccgcggtg gcggccgctc tagaactagt ggatccgatc
     3661 tgccaattga acataacatg gtagttacat atactagtaa tatggttcgg cacacattaa
     3721 aagtataaaa actatctgaa ttacgaatta catatattgg tcataaaaat caatcaatca
     3781 tcgtgtgttt tatatgtctc ttatctaagt ataagaatat ccatagttaa tattcactta
     3841 cgctaccttt taacctgtaa tcattgtcaa caggatatgt tacgacccac attgataaac
     3901 gctagtattt ctttttcctc ttcttattgg ccggctgtct ctatactccc ctatagtctg
     3961 tttcttttcg tttcgattgt tttacgtttg aggcctcgtg gcgcacatgg tacgctgtgg
     4021 tgctcgcggc tgggaacgaa actctgggag ctgcgattgg cagaattccg gctagaacta
     4081 gtggatcccc cggcgcccgg cggctcgagc ccgggcagct tgcatgccgc cgggcgccat
     4141 gcctgcaggt cgactctaga aaacatgagg atcacccatg tctgcaggtc gactctagaa
     4201 aacatgagga tcacccatgt ctgcaggtcg actctagagg atcggaattc ccccatatcc
     4261 aacttccaat ttaatctttc ttttttaatt ttcacttatt tgcgatacag aaagaaaaaa
     4321 gcgatagtaa ctattgaatt ttgtttggat ttggttagat tagatatggt ttctctttat
     4381 atttacatgc taaaaatggg ctacaccaga gatacataat tagatatata tacgccagta
     4441 caccttatcg gcccaagcct tgtcccaagg cagcgttttg ttcttggaaa cgctgcccta
     4501 cacgttcgct atgcttcaag aacttttctg agcacttcat gatgcatgtt tgttccttat
     4561 tggttagctt tgatgttgtg aagtcattga cacgtaccca attcgcccta tagtgagtcg
     4621 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     4681 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     4741 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcgacgcg
     4801 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4861 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4921 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4981 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5041 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5101 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5161 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5221 aattttaaca aaatattaac gtttacaatt tcctgatgcg gtattttctc cttacgcatc
     5281 tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt
     5341 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     5401 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     5461 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     5521 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     5581 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     5641 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     5701 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     5761 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     5821 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     5881 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     5941 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     6001 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     6061 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     6121 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     6181 gacatgattt atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact
     6241 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     6301 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaagaaac
     6361 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga attgaaaagc tgtggtatgg
     6421 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     6481 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     6541 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     6601 a
//
