LOCUS       sequence-249313-        6458 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 6458)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6458)
  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-468    N-Tag UnaG[2] L32 FKBP mCherry
FEATURES             Location/Qualifiers
     source          1..6458
                     /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"
     CDS             1289..1612
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(1626..2200,2201..2203,2204..2212)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1793..1810)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2020..2039
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2213..2923
                     /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(2358..2376)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2626..2645
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2969..3101
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3110..3698
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3163..3183)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3581..3592)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3725..3742)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3731..3842
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3938..3958)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3938..3956)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3977..3993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3977..3993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3990..4012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4095..4114)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4160..4215
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4214..4233)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4594..5182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4674..4693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5353..6144,6145..6213))
                     /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     5976..5995
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6214..6318)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6386..6404
                     /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 caccatgggg gtgaagtctg tggtgaatct ggtaggcgag aaactcgtgt acgtccagaa
     1081 gtgggatggg aaggaaacca cgtatgtgcg agagatcaag gacgggaagt tggttgtcac
     1141 cctgaccatg ggcgatgtgg ttgctgtgcg ctcatatcgg agagccactg aagacgtggg
     1201 tggcggaggg agcgaaggcg gcggatccgg agggcctggc tctggcggag aggggagcgc
     1261 tggaggcggg tcagccggtg gtgggagtgg agtgcaggtg gaaaccatct ccccaggaga
     1321 cgggcgcacc ttccccaagc gcggccagac ctgcgtggtg cactacaccg ggatgcttga
     1381 agatggaaag aaatttgatt cctcccggga cagaaacaag ccctttaagt ttatgctagg
     1441 caagcaggag gtgatccgag gctgggaaga aggggttgcc cagatgagtg tgggtcagag
     1501 agccaaactg actatatctc cagattatgc ctatggtgcc actgggcacc caggcatcat
     1561 cccaccacat gccactctcg tcttcgatgt ggagcttcta aaactggaat aagaattcag
     1621 atccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     1681 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     1741 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     1801 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctggt agcttcttga
     1861 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     1921 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     1981 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     2041 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     2101 cggtacacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     2161 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag
     2221 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     2281 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     2341 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     2401 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     2461 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     2521 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     2581 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     2641 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     2701 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     2761 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     2821 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     2881 gggccgccac tccaccggcg gcatggacga gctgtacaag taaaccggtg ctcgaggtac
     2941 cgatgaatag ctaaggtcga ggccgcaggt aagtatcaag gttacaagac aggtttaagg
     3001 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     3061 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtcgaca atcaacctct
     3121 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3181 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3241 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3301 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3361 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3421 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3481 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3541 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3601 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3661 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacga
     3721 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3781 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3841 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3901 ggggaggatt gggaagacaa tagcccagct tttgttccct ttagtgaggg ttaattgcgc
     3961 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact
     4021 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     4081 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     4141 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     4201 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     4261 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     4321 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     4381 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     4441 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     4501 ctactcccag tcatagctgt ccctcttctc ttatggagat cggccggccg ccggcgtcga
     4561 cgcgcgcggc cgcggcgcgc cgttgctggc gtttttccat aggctccgcc cccctgacga
     4621 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4681 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4741 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4801 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4861 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4921 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4981 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5041 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5101 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5161 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5221 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5281 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5341 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5401 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5461 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5521 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5581 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5641 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5701 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5761 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5821 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5881 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5941 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6001 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6061 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6121 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6181 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6241 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6301 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6361 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctacgcgc
     6421 gcgccggcgg cgcgccgtcg acggccggcc gcggccgc
//
