LOCUS       sequence-249263-        6018 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 6018)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6018)
  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-131    N-Tag GFP[1] L20 {Esp3I(2x)} mCherry
FEATURES             Location/Qualifiers
     source          1..6018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        533..736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     687..707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     733..757
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          872..1004
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(1186..1760,1761..1763,1764..1772)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1353..1370)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1580..1599
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1773..2483
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1918..1936)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2186..2205
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2529..2661
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2670..3258
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2723..2743)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3141..3152)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3285..3302)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3291..3402
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3498..3518)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3498..3516)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3537..3553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3537..3553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3550..3572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3655..3674)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3720..3775
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3774..3793)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4154..4742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4234..4253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4913..5704,5705..5773))
                     /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     5536..5555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5774..5878)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5946..5964
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcctgcctcc aggcctcaat attggccatt agccatatta ttcattggtt atatagcata
       61 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
      121 tattggctca tgtccaatat gaccgccatg ttggcattga ttattgacta gttattaata
      181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
      421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
      481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      541 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      661 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
      721 ctatataagc agagctcgtt tagtgaaccg tcagatcact agaagcttta ttgcggtagt
      781 ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct cgaacttaag
      841 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
      901 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      961 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccaggc
     1021 caccatggat ctgcctgatg accactacct gagcacacag accatcctct ccaaggacct
     1081 taacgggagt cctggcggtg gatccggagg ctctgggagc tctgccagcg gtggcagcac
     1141 ctcccgagac gccgaggcac gccgtctcct aagaattcag atccgcccct ctccctcccc
     1201 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     1261 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     1321 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     1381 gaatgtcgtg aaggaagcag ttcctctggt agcttcttga agacaaacaa cgtctgtagc
     1441 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     1501 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     1561 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     1621 ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat gctttacatg
     1681 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     1741 ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag gaggataaca
     1801 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     1861 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     1921 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     1981 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     2041 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     2101 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     2161 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     2221 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     2281 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     2341 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     2401 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     2461 gcatggacga gctgtacaag taaaccggtg ctcgaggtac cgatgaatag ctaaggtcga
     2521 ggccgcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
     2581 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
     2641 actttgcctt tctctccaca ggtgtcgaca atcaacctct ggattacaaa atttgtgaaa
     2701 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2761 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2821 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2881 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2941 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3001 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3061 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3121 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3181 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3241 tttgggccgc ctccccgcct ggaattcgag ctcggtacga tcagcctcga ctgtgccttc
     3301 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3361 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3421 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3481 tagcccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     3541 tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact cctcaggtgc aggctgccta
     3601 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     3661 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     3721 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3781 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     3841 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     3901 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     3961 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     4021 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     4081 ccctcttctc ttatggagat cggccggccg ccggcgtcga cgcgcgcggc cgcggcgcgc
     4141 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4201 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4261 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4321 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4381 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4441 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4501 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4561 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4621 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4681 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4741 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4801 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4861 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4921 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4981 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5041 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5101 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5161 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5221 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5281 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5341 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5401 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5461 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5521 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5581 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5641 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5701 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5761 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5821 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5881 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5941 ataaaaatag gcgtatcacg aggccctttc gtctacgcgc gcgccggcgg cgcgccgtcg
     6001 acggccggcc gcggccgc
//
