LOCUS       sequence-249325-        6506 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 6506)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6506)
  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-431    N-Tag Venus173/155[2] L32 FRB mCh...
FEATURES             Location/Qualifiers
     source          1..6506
                     /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             1382..1657
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRIS"
     misc_feature    join(1674..2248,2249..2251,2252..2260)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1841..1858)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2068..2087
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2261..2971
                     /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(2406..2424)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2674..2693
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          3017..3149
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3158..3746
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3211..3231)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3629..3640)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3773..3790)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3779..3890
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3986..4006)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3986..4004)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4025..4041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4025..4041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4038..4060)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4143..4162)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4208..4263
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4262..4281)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4642..5230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4722..4741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5401..6192,6193..6261))
                     /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     6024..6043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6262..6366)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6434..6452
                     /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 acgtgggtgg cggagggagc gaaggcggcg gatccggagg
     1321 gcctggctct ggcggagagg ggagcgctgg aggcgggtca gccggtggtg ggagtgtggc
     1381 catcctctgg catgagatgt ggcatgaagg cctggaagag gcatctcgtt tgtactttgg
     1441 ggaaaggaac gtgaaaggca tgtttgaggt gctggagccc ttgcatgcta tgatggaacg
     1501 gggcccccag actctgaagg aaacatcctt taatcaggcc tatggtcgag atttaatgga
     1561 ggcccaagag tggtgcagga agtacatgaa atcagggaat gtcaaggacc tcacccaagc
     1621 ctgggacctc tattatcatg tgttccgacg aatctcataa gaattcagat ccgcccctct
     1681 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     1741 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     1801 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     1861 ggtctgttga atgtcgtgaa ggaagcagtt cctctggtag cttcttgaag acaaacaacg
     1921 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     1981 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     2041 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     2101 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtacacatgc
     2161 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     2221 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga
     2281 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     2341 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     2401 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     2461 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     2521 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     2581 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     2641 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     2701 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     2761 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     2821 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     2881 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     2941 caccggcggc atggacgagc tgtacaagta aaccggtgct cgaggtaccg atgaatagct
     3001 aaggtcgagg ccgcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
     3061 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
     3121 tgacatccac tttgcctttc tctccacagg tgtcgacaat caacctctgg attacaaaat
     3181 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3241 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3301 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3361 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3421 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3481 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3541 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     3601 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3661 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3721 gatctccctt tgggccgcct ccccgcctgg aattcgagct cggtacgatc agcctcgact
     3781 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3841 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3901 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3961 gaagacaata gcccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     4021 catggtcata gctgtttcct gtgtgaaatt gttatccgct aattcactcc tcaggtgcag
     4081 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     4141 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
     4201 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     4261 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
     4321 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
     4381 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     4441 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     4501 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     4561 atagctgtcc ctcttctctt atggagatcg gccggccgcc ggcgtcgacg cgcgcggccg
     4621 cggcgcgccg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4801 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5041 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5101 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5161 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5341 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5401 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5461 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5521 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5581 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5641 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5701 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5761 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5821 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5881 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5941 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6001 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6061 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6121 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6181 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6241 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6301 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6361 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     6421 attaacctat aaaaataggc gtatcacgag gccctttcgt ctacgcgcgc gccggcggcg
     6481 cgccgtcgac ggccggccgc ggccgc
//
