LOCUS       sequence-249261-        5988 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 5988)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5988)
  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-294    N-Tag GFP[1] L10 {Esp3I(2x)} mCherry
FEATURES             Location/Qualifiers
     source          1..5988
                     /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(1156..1730,1731..1733,1734..1742)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1323..1340)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1550..1569
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1743..2453
                     /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(1888..1906)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2156..2175
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2499..2631
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2640..3228
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2693..2713)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3111..3122)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3255..3272)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3261..3372
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3468..3488)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3468..3486)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3507..3523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3507..3523)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3520..3542)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3625..3644)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3690..3745
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3744..3763)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4124..4712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4204..4223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4883..5674,5675..5743))
                     /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     5506..5525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5744..5848)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5916..5934
                     /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 taacgggagc tctgccagcg gtggcagcac ctcccgagac gccgaggcac gccgtctcct
     1141 aagaattcag atccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt
     1201 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     1261 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     1321 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctggt
     1381 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     1441 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     1501 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     1561 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     1621 tctggggcct cggtacacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     1681 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa
     1741 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     1801 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1861 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     1921 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     1981 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     2041 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     2101 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     2161 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2221 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2281 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2341 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2401 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taaaccggtg
     2461 ctcgaggtac cgatgaatag ctaaggtcga ggccgcaggt aagtatcaag gttacaagac
     2521 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     2581 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtcgaca
     2641 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2701 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2761 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2821 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2881 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2941 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3001 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     3061 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3121 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3181 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag
     3241 ctcggtacga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3301 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3361 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3421 ggacagcaag ggggaggatt gggaagacaa tagcccagct tttgttccct ttagtgaggg
     3481 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3541 ctaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     3601 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     3661 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     3721 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     3781 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     3841 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     3901 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     3961 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     4021 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat cggccggccg
     4081 ccggcgtcga cgcgcgcggc cgcggcgcgc cgttgctggc gtttttccat aggctccgcc
     4141 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4201 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4261 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4321 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4381 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4441 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4501 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4561 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4621 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4681 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4741 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4801 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4861 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4921 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4981 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5041 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5101 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5161 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5221 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5281 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5341 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5401 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5461 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5521 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5581 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5641 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5701 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5761 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5821 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5881 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5941 gtctacgcgc gcgccggcgg cgcgccgtcg acggccggcc gcggccgc
//
