LOCUS       sequence-249299-        6383 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 6383)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6383)
  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-478    N-Tag UnaG[2] L20 FRB mCherry
FEATURES             Location/Qualifiers
     source          1..6383
                     /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             1259..1534
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRIS"
     misc_feature    join(1551..2125,2126..2128,2129..2137)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1718..1735)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1945..1964
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2138..2848
                     /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(2283..2301)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2551..2570
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2894..3026
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3035..3623
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3088..3108)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3506..3517)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3650..3667)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3656..3767
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3863..3883)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3863..3881)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3902..3918)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3902..3918)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3915..3937)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4020..4039)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4085..4140
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4139..4158)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4519..5107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4599..4618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5278..6069,6070..6138))
                     /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     5901..5920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6139..6243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6311..6329
                     /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 aagggagtcc
     1201 tggcggtgga tccggaggct ctgggagctc tgccagcggt ggcagcacct ccgtggccat
     1261 cctctggcat gagatgtggc atgaaggcct ggaagaggca tctcgtttgt actttgggga
     1321 aaggaacgtg aaaggcatgt ttgaggtgct ggagcccttg catgctatga tggaacgggg
     1381 cccccagact ctgaaggaaa catcctttaa tcaggcctat ggtcgagatt taatggaggc
     1441 ccaagagtgg tgcaggaagt acatgaaatc agggaatgtc aaggacctca cccaagcctg
     1501 ggacctctat tatcatgtgt tccgacgaat ctcataagaa ttcagatccg cccctctccc
     1561 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     1621 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     1681 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     1741 ctgttgaatg tcgtgaagga agcagttcct ctggtagctt cttgaagaca aacaacgtct
     1801 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     1861 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     1921 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     1981 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggta cacatgcttt
     2041 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2101 ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagga
     2161 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     2221 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     2281 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     2341 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2401 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2461 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2521 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     2581 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     2641 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     2701 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     2761 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     2821 cggcggcatg gacgagctgt acaagtaaac cggtgctcga ggtaccgatg aatagctaag
     2881 gtcgaggccg caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact
     2941 gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga
     3001 catccacttt gcctttctct ccacaggtgt cgacaatcaa cctctggatt acaaaatttg
     3061 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3121 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3181 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3241 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3301 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3361 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3421 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3481 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3541 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3601 ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacgatcagc ctcgactgtg
     3661 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3721 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3781 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3841 gacaatagcc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     3901 ggtcatagct gtttcctgtg tgaaattgtt atccgctaat tcactcctca ggtgcaggct
     3961 gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat
     4021 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     4081 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     4141 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     4201 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
     4261 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     4321 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     4381 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     4441 gctgtccctc ttctcttatg gagatcggcc ggccgccggc gtcgacgcgc gcggccgcgg
     4501 cgcgccgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4561 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4621 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4681 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4741 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4801 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4861 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4921 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4981 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5041 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5101 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5161 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5221 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5281 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5341 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5401 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5461 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5521 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5581 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5641 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5701 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5761 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5821 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5881 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5941 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6001 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6061 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6121 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6241 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     6301 aacctataaa aataggcgta tcacgaggcc ctttcgtcta cgcgcgcgcc ggcggcgcgc
     6361 cgtcgacggc cggccgcggc cgc
//
