LOCUS       sequence-249507-        5606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PPI (BiFC/TriFC) assay vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5606)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5606)
  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"
COMMENT     Sequence Label: pRDAX-019    FLAG IFP1.4[1] IRES IFP1.4[2]
FEATURES             Location/Qualifiers
     source          1..5606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        173..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      747..756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             756..779
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(1195..1769,1770..1772,1773..1781)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1362..1379)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1589..1608
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2358..2381
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2506..2627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2543..2562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2597..2616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2634..3089)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2771..2792)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2983..3002
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3116..3220
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3222..3579
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3222..3413
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3245..3265)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3430..3565
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3614..4408
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3614..4408
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3668..3687)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4278..4297
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4596..4615)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4640..4687
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5016..5604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5505..5524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
       61 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
      721 agatctcgag ctcaagcttc gaattcgcca ccatggacta caaagacgat gacgataaag
      781 gaagtgctcg ggatccattg cctttcttcc cacccttgta cctcggtgga ccagagatca
      841 ccaccgagaa ctgcgagagg gaaccgatac acattcccgg gtccattcag ccccatggtg
      901 cgctgctgac tgctgacgga cattcagggg aagtgctgca agtcagcctg aatgcagcaa
      961 cctttctggg ccaagaaccc actgtgcttc gcgggcagac cttggctgct ctgctgcctg
     1021 agcagtggcc agccctccaa gcagctcttc cacccggttg tccagacgcc ctccagtatc
     1081 gtgccactct ggactggcct gctgccggcc accttagcct gacagtgcac agggtagctg
     1141 agctgctcat cctcgaattc gagcccaccg aggcatggta agaattcaga tccgcccctc
     1201 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     1261 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     1321 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     1381 aggtctgttg aatgtcgtga aggaagcagt tcctctggta gcttcttgaa gacaaacaac
     1441 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     1501 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     1561 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     1621 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtacacatg
     1681 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     1741 ggttttcctt tgaaaaacac gatgataata tggccacaac catggacagc attggacccc
     1801 atgcgctgag gaacgctatg ttcgcccttg aatctgctcc caatctccga gcactggctg
     1861 aagtcgccac acagacagtc agggagctga caggctttga tcgggtgatg ctgtacaagt
     1921 ttgccccaga tgcaacgggt gagatgatcg cagaagccag acgggaaggg atgcaggcct
     1981 tcctgggcca cagatttccg gcctcacata cgcctgctca ggccagagca ctgtacaccc
     2041 gacacctcct gagactgact gcggacacta gagcagctgc cgtgccactc gatcccgtgc
     2101 tcaacccaca gaccaatgcg cctacccctt tgggaggagc cgtactgagg gccacatccc
     2161 ctatgcacat gcagtatctg cgcaacatgg gcgttgggag tagtctgtct gtgtccgttg
     2221 tcgttggagg gcaactgtgg ggcttgatcg tctgccacca tcagacaccg tatgtgcttc
     2281 ctcctgactt gcgtacaact ctggaggagc tgggccgcaa gcttagcggc caggtgcagc
     2341 ggaaagaggc cactggtgat tacaaggatg acgatgacaa gtaagcggcc gcgactctag
     2401 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2461 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2521 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2581 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     2641 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2701 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2761 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2821 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2881 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2941 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3001 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3061 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3121 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3181 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3241 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3301 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3361 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3421 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3481 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3541 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3601 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3661 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3721 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3781 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3841 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3901 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3961 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4021 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4081 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4141 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4201 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4261 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4321 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4381 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4441 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4501 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4561 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4621 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4681 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4741 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4801 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4861 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4921 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4981 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5041 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5101 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5161 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5221 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5281 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5341 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5401 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5461 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5521 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5581 tcaggggggc ggagcctatg gaaaaa
//
