LOCUS       sequence-249335-        6509 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 6509)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6509)
  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-310    N-Tag Venus173/155[2] L20 FKBP mC...
FEATURES             Location/Qualifiers
     source          1..6509
                     /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             1340..1663
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(1677..2251,2252..2254,2255..2263)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1844..1861)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2071..2090
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2264..2974
                     /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(2409..2427)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2677..2696
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          3020..3152
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3161..3749
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3214..3234)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3632..3643)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3776..3793)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3782..3893
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3989..4009)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3989..4007)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4028..4044)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4028..4044)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4041..4063)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4146..4165)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4211..4266
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4265..4284)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4645..5233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4725..4744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5404..6195,6196..6264))
                     /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     6027..6046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6265..6369)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6437..6455
                     /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 aagcagaaga atgggattaa ggcgaacttc aagatccgac acaacatcga
     1081 ggatggtggg gtccagcttg ccgaccatta tcagcagaac acacccattg gggatggacc
     1141 agtcctgttg ccggataacc actacctcag ttaccagagc aaactgtcca aagatcccaa
     1201 tgagaagagg gaccatatgg tactgctcga attcgtgact gcagcaggca ttacccttgg
     1261 gatggatgaa ctttataagg ggagtcctgg cggtggatcc ggaggctctg ggagctctgc
     1321 cagcggtggc agcacctccg gagtgcaggt ggaaaccatc tccccaggag acgggcgcac
     1381 cttccccaag cgcggccaga cctgcgtggt gcactacacc gggatgcttg aagatggaaa
     1441 gaaatttgat tcctcccggg acagaaacaa gccctttaag tttatgctag gcaagcagga
     1501 ggtgatccga ggctgggaag aaggggttgc ccagatgagt gtgggtcaga gagccaaact
     1561 gactatatct ccagattatg cctatggtgc cactgggcac ccaggcatca tcccaccaca
     1621 tgccactctc gtcttcgatg tggagcttct aaaactggaa taagaattca gatccgcccc
     1681 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     1741 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     1801 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     1861 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg tagcttcttg aagacaaaca
     1921 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     1981 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     2041 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     2101 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtacaca
     2161 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     2221 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga
     2281 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     2341 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     2401 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     2461 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     2521 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     2581 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     2641 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     2701 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     2761 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     2821 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     2881 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     2941 ctccaccggc ggcatggacg agctgtacaa gtaaaccggt gctcgaggta ccgatgaata
     3001 gctaaggtcg aggccgcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
     3061 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
     3121 tactgacatc cactttgcct ttctctccac aggtgtcgac aatcaacctc tggattacaa
     3181 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3241 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3301 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3361 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3421 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3481 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3541 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3601 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3661 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3721 tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacg atcagcctcg
     3781 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3841 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3901 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3961 tgggaagaca atagcccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     4021 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctaattcac tcctcaggtg
     4081 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     4141 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     4201 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     4261 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     4321 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     4381 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     4441 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     4501 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     4561 gtcatagctg tccctcttct cttatggaga tcggccggcc gccggcgtcg acgcgcgcgg
     4621 ccgcggcgcg ccgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5161 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5221 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5281 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5341 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5401 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5461 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5521 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5581 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5641 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5701 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5761 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5821 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5881 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5941 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6001 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6061 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6121 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6181 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6241 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6301 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6361 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     6421 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctacgcg cgcgccggcg
     6481 gcgcgccgtc gacggccggc cgcggccgc
//
