LOCUS       sequence-249264-        6051 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 6051)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6051)
  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-132    C-Tag GFP[1] L32 {Esp3I(2x)} mCherry
FEATURES             Location/Qualifiers
     source          1..6051
                     /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"
     misc_feature    join(1219..1793,1794..1796,1797..1805)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1386..1403)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1613..1632
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1806..2516
                     /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(1951..1969)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2219..2238
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2562..2694
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2703..3291
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2756..2776)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3174..3185)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3318..3335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3324..3435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3531..3551)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3531..3549)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3570..3586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3570..3586)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3583..3605)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3688..3707)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3753..3808
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3807..3826)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4187..4775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4267..4286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4946..5737,5738..5806))
                     /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     5569..5588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5807..5911)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5979..5997
                     /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 cacccgagac gccgaggcac gccgtctccg acgtgggtgg cggagggagc gaaggcggcg
     1081 gatccggagg gcctggctct ggcggagagg ggagcgctgg aggcgggtca gccggtggtg
     1141 ggagtgatct gcctgatgac cactacctga gcacacagac catcctctcc aaggacctta
     1201 actaagaatt cagatccgcc cctctccctc ccccccccct aacgttactg gccgaagccg
     1261 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     1321 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     1381 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     1441 ggtagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     1501 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     1561 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     1621 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     1681 tgatctgggg cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     1741 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     1801 caaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     1861 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     1921 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     1981 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     2041 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     2101 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     2161 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     2221 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     2281 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     2341 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     2401 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2461 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaaaccg
     2521 gtgctcgagg taccgatgaa tagctaaggt cgaggccgca ggtaagtatc aaggttacaa
     2581 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
     2641 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcg
     2701 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2761 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2821 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2881 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2941 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3001 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3061 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     3121 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3181 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3241 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaattc
     3301 gagctcggta cgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3361 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3421 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3481 gcaggacagc aagggggagg attgggaaga caatagccca gcttttgttc cctttagtga
     3541 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3601 ccgctaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
     3661 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
     3721 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     3781 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     3841 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     3901 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     3961 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     4021 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     4081 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga gatcggccgg
     4141 ccgccggcgt cgacgcgcgc ggccgcggcg cgccgttgct ggcgtttttc cataggctcc
     4201 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4261 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4321 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4381 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4441 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4501 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4561 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4621 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4681 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4741 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4801 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4861 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4921 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4981 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5041 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5101 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5161 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5221 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5281 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5341 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5401 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5461 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5521 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5581 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5641 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5701 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5761 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5821 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5881 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5941 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6001 ttcgtctacg cgcgcgccgg cggcgcgccg tcgacggccg gccgcggccg c
//
