LOCUS       sequence-249332-        6479 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 6479)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6479)
  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-307    C-Tag Venus173/155[2] L10 FKBP mC...
FEATURES             Location/Qualifiers
     source          1..6479
                     /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             join(1025..1027,1028..1348)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     misc_feature    join(1647..2221,2222..2224,2225..2233)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1814..1831)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2041..2060
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2234..2944
                     /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(2379..2397)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2647..2666
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2990..3122
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3131..3719
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3184..3204)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3602..3613)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3746..3763)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3752..3863
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3959..3979)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3959..3977)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3998..4014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3998..4014)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4011..4033)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4116..4135)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4181..4236
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4235..4254)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4615..5203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4695..4714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5374..6165,6166..6234))
                     /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     5997..6016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6235..6339)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6407..6425
                     /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 caccatggga gtgcaggtgg aaaccatctc cccaggagac gggcgcacct tccccaagcg
     1081 cggccagacc tgcgtggtgc actacaccgg gatgcttgaa gatggaaaga aatttgattc
     1141 ctcccgggac agaaacaagc cctttaagtt tatgctaggc aagcaggagg tgatccgagg
     1201 ctgggaagaa ggggttgccc agatgagtgt gggtcagaga gccaaactga ctatatctcc
     1261 agattatgcc tatggtgcca ctgggcaccc aggcatcatc ccaccacatg ccactctcgt
     1321 cttcgatgtg gagcttctaa aactggaagg gagctctgcc agcggtggca gcaccagtga
     1381 taagcagaag aatgggatta aggcgaactt caagatccga cacaacatcg aggatggtgg
     1441 ggtccagctt gccgaccatt atcagcagaa cacacccatt ggggatggac cagtcctgtt
     1501 gccggataac cactacctca gttaccagag caaactgtcc aaagatccca atgagaagag
     1561 ggaccatatg gtactgctcg aattcgtgac tgcagcaggc attacccttg ggatggatga
     1621 actttataag taagaattca gatccgcccc tctccctccc ccccccctaa cgttactggc
     1681 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     1741 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     1801 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     1861 gttcctctgg tagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     1921 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     1981 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     2041 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     2101 atgggatctg atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa
     2161 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     2221 tatggccaca accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     2281 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     2341 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     2401 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     2461 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     2521 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     2581 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     2641 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     2701 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     2761 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     2821 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     2881 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     2941 gtaaaccggt gctcgaggta ccgatgaata gctaaggtcg aggccgcagg taagtatcaa
     3001 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     3061 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     3121 aggtgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3181 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3241 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3301 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3361 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3421 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3481 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3541 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3601 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3661 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3721 tggaattcga gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3781 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3841 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3901 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcccagc ttttgttccc
     3961 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4021 attgttatcc gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     4081 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     4141 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     4201 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     4261 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     4321 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     4381 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     4441 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     4501 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     4561 tcggccggcc gccggcgtcg acgcgcgcgg ccgcggcgcg ccgttgctgg cgtttttcca
     4621 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4681 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4741 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4801 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4861 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4921 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4981 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5041 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5101 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5161 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5221 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5281 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5341 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5401 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5461 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5521 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5581 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5641 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5701 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5761 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5821 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5881 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5941 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6001 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6061 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6121 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6181 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6241 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6301 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6361 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6421 gaggcccttt cgtctacgcg cgcgccggcg gcgcgccgtc gacggccggc cgcggccgc
//
