LOCUS       sequence-351667-        7502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector to insert genes of interest having a 
            N-terminal DHFR F[1,2] fusion for DHFR-PCA/BFG-PCA assays..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7502)
  AUTHORS   Evans-Yamamoto D, Rouleau FD, Nanda P, Makanae K, Liu Y, Despres PC,
            Matsuo H, Seki M, Dube AK, Ascencio D, Yachie N, Landry CR
  TITLE     Barcode fusion genetics-protein-fragment complementation assay 
            (BFG-PCA): tools and resources that expand the potential for binary 
            protein interaction discovery.
  JOURNAL   Nucleic Acids Res. 2022 Feb 7. pii: 6523796. doi: 
            10.1093/nar/gkac045.
  PUBMED    35137167
REFERENCE   2  (bases 1 to 7502)
  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. 2022 Feb 7. pii: 6523796. doi: 10.1093/nar/gkac045."
COMMENT     Sequence Label: pDEST-DHFR F[1,2]-N (TRP1)
FEATURES             Location/Qualifiers
     source          1..7502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      109..356
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(125..143)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        1166..1870
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     protein_bind    2223..2347
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2494..2596
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2597..3277
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(2663..2682)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3597..3902
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3781..3800
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3943..4067)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      4326..4513
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4381..4405)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4665..4684)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4666..4684)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4691..4708)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4691..4707)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4700..4722)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4848..5303
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4935..4954)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5145..5166
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5406..6080)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(6081..6361)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6382..6401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6501..6523
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6561..6579)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6620..7123
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7160..7264
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7483..7502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtaccaagat ggcctttggt gggttgaaga aggaaaaaga cagaaacgac ttaattacct
       61 acttgaaaaa agcctgtgag taaacaggcc ccttttcctt tgtcgatatc atgtaattag
      121 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
      181 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
      241 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
      301 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc
      361 tcgggatccg ggccccccct cgagatccgg gatcgaagaa atgatggtaa atgaaatagg
      421 aaatcaagga gcatgaaggc aaaagacaaa tataagggtc gaacgaaaaa taaagtgaaa
      481 agtgttgata tgatgtattt ggctttgcgg cgccgaaaaa acgagtttac gcaattgcac
      541 aatcatgctg actctgtggc ggacccgcgc tcttgccggc ccggcgataa cgctgggcgt
      601 gaggctgtgc ccggcggagt tttttgcgcc tgcattttcc aaggtttacc ctgcgctaag
      661 gggcgagatt ggagaagcaa taagaatgcc ggttggggtt gcgatgatga cgaccacgac
      721 aactggtgtc attatttaag ttgccgaaag aacctgagtg catttgcaac atgagtatac
      781 tagaagaatg agccaagact tgcgagacgc gagtttgccg gtggtgcgaa caatagagcg
      841 accatgacct tgaaggtgag acgcgcataa ccgctagagt actttgaaga ggaaacagca
      901 atagggttgc taccagtata aatagacagg tacatacaac actggaaatg gttgtctgtt
      961 tgagtacgct ttcaattcat ttgggtgtgc actttattat gttacaatat ggaagggaac
     1021 tttacacttc tcctatgcac atatattaat gtaactataa cggtcctaag gtagcgaaat
     1081 taaagtccaa tgctagtaga gaaggggggt aacacccctc cgcgctcttt tccgattttt
     1141 ttctaaaccg tggaatattt cggatatcct tttgttgttt ccgggtgtac aatatggact
     1201 tcctcttttc tggcaaccaa acccatacat cgggattcct ataatacctt cgttggtctc
     1261 cctaacatgt aggtggcgga ggggagatat acaatagaac agataccaga caagacataa
     1321 tgggctaaac aagactacac caattacact gcctcattga tggtggtaca taacgaacta
     1381 atactgtagc cctagacttg atagccatca tcatatcgaa gtttcactac cctttttcca
     1441 tttgccatct attgaagtaa taataggcgc atgcaacttc ttttcttttt ttttcttttc
     1501 tctctccccc gttgttgtct caccatatcc gcaatgacaa aaaaatgatg gaagacacta
     1561 aaggaaaaaa ttaacgacaa agacagcacc aacagatgtc gttgttccag agctgatgag
     1621 gggtatctcg aagcacacga aactttttcc ttccttcatt cacgcacact actctctaat
     1681 gagcaacggt atacggcctt ccttccagtt acttgaattt gaaataaaaa aaagtttgct
     1741 gtcttgctat caagtataaa tagacctgca attattaatc ttttgtttcc tcgtcattgt
     1801 tctcgttccc tttcttcctt gtttcttttt ctgcacaata tttcaagcta taccaagcat
     1861 acaatcaact ccaagcttgg atggttcgac cattgaactg catcgtcgcc gtgtcccaaa
     1921 atatggggat tggcaagaac ggagactatc cctggcctcc gctcaggaac gagtccaagt
     1981 acttccaaag aatgaccaca acctcttcag tggaaggtaa acagaatctg gtgattatgg
     2041 gtaggaaaac ctggttctcc attcctgaga agaatcgacc tttaaaggac agaattaata
     2101 tagttctcag tagagaactc aaagaaccac cacgaggagc tcattttctt gccaaaagtt
     2161 tggatgatgc cttaagactt attgaacaac cggaattggg taccggagga ggaagtacta
     2221 gtacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat
     2281 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
     2341 cactatggcg gccgctaagt tggcagcatc accctacgca ctttgcgccg aataaatacc
     2401 tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg ataccgggaa
     2461 gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag gttccaactt
     2521 tcaccataat gaaataagat cactaccggg cgtatttttt gagtcatcga gattttcagg
     2581 agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca
     2641 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     2701 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     2761 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat
     2821 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     2881 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     2941 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     3001 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
     3061 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     3121 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     3181 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     3241 gcaggcgggg cgtaatctag aggatccggc ttactaaaag ccagataaca gtatgcgtat
     3301 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
     3361 aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
     3421 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
     3481 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
     3541 cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc
     3601 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
     3661 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
     3721 tgtcagataa agtcccccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
     3781 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
     3841 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
     3901 aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt
     3961 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
     4021 tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggtttg atggccgcta
     4081 agtaagtaag agctttaagt aagtaatggc aaacagctat tatgggtatt atgggttagg
     4141 gataacaggg taataagctc taagtaagta acggccgcca ccgcggtgga gctttggact
     4201 tcttcgccag aggtttggtc aagtctccaa tcaaggttgt cggcttgtct accttgccag
     4261 aaatttacga aaagatggaa aagggtcaaa tcgttggtag atacgttgtt gacacttcta
     4321 aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa
     4381 gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta
     4441 actctttcct gtaggtcagg ttgctttctc aggtatagca tgaggtcgct cttattgacc
     4501 acacctctac cggcatgccg agcaaatgcc tgcaaatcgc tccccatttc acccaattgt
     4561 agatatgcta actccagcaa tgagttgatg aatctcggtg tgtattttat gtcctcagag
     4621 gacaacacct gttgtaatcg ttcttccaca cggatcccaa ttcgccctat agtgagtcgt
     4681 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4741 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4801 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     4861 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4921 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4981 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5041 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5101 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5161 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5221 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5281 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     5341 atttcacacc gcatagatcg gcaagtgcac aaacaatact taaataaata ctactcagta
     5401 ataacctatt tcttagcatt tttgacgaaa tttgctattt tgttagagtc ttttacacca
     5461 tttgtctcca cacctccgct tacatcaaca ccaataacgc catttaatct aagcgcatca
     5521 ccaacatttt ctggcgtcag tccaccagct aacataaaat gtaagctttc ggggctctct
     5581 tgccttccaa cccagtcaga aatcgagttc caatccaaaa gttcacctgt cccacctgct
     5641 tctgaatcaa acaagggaat aaacgaatga ggtttctgtg aagctgcact gagtagtatg
     5701 ttgcagtctt ttggaaatac gagtctttta ataactggca aaccgaggaa ctcttggtat
     5761 tcttgccacg actcatctcc atgcagttgg acgatatcaa tgccgtaatc attgaccaga
     5821 gccaaaacat cctccttagg ttgattacga aacacgccaa ccaagtattt cggagtgcct
     5881 gaactatttt tatatgcttt tacaagactt gaaattttcc ttgcaataac cgggtcaatt
     5941 gttctctttc tattgggcac acatataata cccagcaagt cagcatcgga atctagagca
     6001 cattctgcgg cctctgtgct ctgcaagccg caaactttca ccaatggacc agaactacct
     6061 gtgaaattaa taacagacat actccaagct gcctttgtgt gcttaatcac gtatactcac
     6121 gtgctcaata gtcaccaatg ccctccctct tggccctctc cttttctttt ttcgaccgaa
     6181 ttaattctta atcggcaaaa aaagaaaagc tccggatcaa gattgtacgt aaggtgacaa
     6241 gctatttttc aataaagaat atcttccact actgccatct ggcgtcataa ctgcaaagta
     6301 cacatatatt acgatgctgt tctattaaat gcttcctata ttatatatat agtaatgtcg
     6361 tgatctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     6421 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     6481 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     6541 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     6601 aataatggtt tcttagacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa
     6661 tgatggaata atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat
     6721 tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa
     6781 ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc
     6841 agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg
     6901 tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa
     6961 gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa
     7021 aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat ttatatattt
     7081 atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg
     7141 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     7201 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     7261 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7321 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     7381 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     7441 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7501 at
//
