LOCUS       sequence-249297-        6353 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 6353)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6353)
  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-476    N-Tag UnaG[2] L10 FRB mCherry
FEATURES             Location/Qualifiers
     source          1..6353
                     /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             1229..1504
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRIS"
     misc_feature    join(1521..2095,2096..2098,2099..2107)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1688..1705)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1915..1934
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2108..2818
                     /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(2253..2271)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2521..2540
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2864..2996
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3005..3593
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3058..3078)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3476..3487)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3620..3637)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3626..3737
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3833..3853)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3833..3851)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3872..3888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3872..3888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3885..3907)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3990..4009)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4055..4110
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4109..4128)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4489..5077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4569..4588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5248..6039,6040..6108))
                     /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     5871..5890
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6109..6213)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6281..6299
                     /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 caccatgggg gtgaagtctg tggtgaatct ggtaggcgag aaactcgtgt acgtccagaa
     1081 gtgggatggg aaggaaacca cgtatgtgcg agagatcaag gacgggaagt tggttgtcac
     1141 cctgaccatg ggcgatgtgg ttgctgtgcg ctcatatcgg agagccactg aagggagctc
     1201 tgccagcggt ggcagcacct ccgtggccat cctctggcat gagatgtggc atgaaggcct
     1261 ggaagaggca tctcgtttgt actttgggga aaggaacgtg aaaggcatgt ttgaggtgct
     1321 ggagcccttg catgctatga tggaacgggg cccccagact ctgaaggaaa catcctttaa
     1381 tcaggcctat ggtcgagatt taatggaggc ccaagagtgg tgcaggaagt acatgaaatc
     1441 agggaatgtc aaggacctca cccaagcctg ggacctctat tatcatgtgt tccgacgaat
     1501 ctcataagaa ttcagatccg cccctctccc tccccccccc ctaacgttac tggccgaagc
     1561 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     1621 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     1681 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     1741 ctggtagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     1801 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     1861 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     1921 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     1981 tctgatctgg ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     2041 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     2101 cacaaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     2161 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     2221 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     2281 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2341 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2401 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2461 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2521 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     2581 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     2641 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     2701 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     2761 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaaac
     2821 cggtgctcga ggtaccgatg aatagctaag gtcgaggccg caggtaagta tcaaggttac
     2881 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
     2941 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
     3001 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3061 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3121 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3181 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3241 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3301 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3361 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     3421 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3481 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3541 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     3601 tcgagctcgg tacgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3661 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3721 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3781 gggcaggaca gcaaggggga ggattgggaa gacaatagcc cagcttttgt tccctttagt
     3841 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3901 atccgctaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     3961 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     4021 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     4081 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     4141 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     4201 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     4261 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     4321 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     4381 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatcggcc
     4441 ggccgccggc gtcgacgcgc gcggccgcgg cgcgccgttg ctggcgtttt tccataggct
     4501 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4561 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4621 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4681 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4741 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4801 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4861 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4921 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4981 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5041 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5101 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5161 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5221 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5281 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5341 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5401 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5461 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5521 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5581 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5641 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5701 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5761 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5821 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5881 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5941 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6001 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6061 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6121 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6181 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6241 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6301 ctttcgtcta cgcgcgcgcc ggcggcgcgc cgtcgacggc cggccgcggc cgc
//
