LOCUS       sequence-249309-        6392 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 6392)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6392)
  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-464    N-Tag UnaG[2] L10 FKBP mCherry
FEATURES             Location/Qualifiers
     source          1..6392
                     /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             1223..1546
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(1560..2134,2135..2137,2138..2146)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1727..1744)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1954..1973
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2147..2857
                     /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(2292..2310)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2560..2579
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2903..3035
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3044..3632
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3097..3117)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3515..3526)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3659..3676)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3665..3776
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3872..3892)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3872..3890)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3911..3927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3911..3927)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3924..3946)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4029..4048)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4094..4149
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4148..4167)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4528..5116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4608..4627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5287..6078,6079..6147))
                     /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     5910..5929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6148..6252)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6320..6338
                     /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 ccggagtgca ggtggaaacc atctccccag gagacgggcg
     1261 caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg
     1321 aaagaaattt gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca
     1381 ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa
     1441 actgactata tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc
     1501 acatgccact ctcgtcttcg atgtggagct tctaaaactg gaataagaat tcagatccgc
     1561 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     1621 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     1681 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     1741 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggtagcttc ttgaagacaa
     1801 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     1861 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     1921 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     1981 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac
     2041 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     2101 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg
     2161 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     2221 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     2281 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     2341 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     2401 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     2461 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     2521 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     2581 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     2641 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     2701 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     2761 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     2821 ccactccacc ggcggcatgg acgagctgta caagtaaacc ggtgctcgag gtaccgatga
     2881 atagctaagg tcgaggccgc aggtaagtat caaggttaca agacaggttt aaggagacca
     2941 atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg
     3001 tcttactgac atccactttg cctttctctc cacaggtgtc gacaatcaac ctctggatta
     3061 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3121 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3181 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3241 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3301 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3361 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3421 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     3481 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3541 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3601 gagtcggatc tccctttggg ccgcctcccc gcctggaatt cgagctcggt acgatcagcc
     3661 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3721 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3781 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3841 gattgggaag acaatagccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     3901 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctaatt cactcctcag
     3961 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     4021 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     4081 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     4141 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     4201 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     4261 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     4321 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     4381 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     4441 ccagtcatag ctgtccctct tctcttatgg agatcggccg gccgccggcg tcgacgcgcg
     4501 cggccgcggc gcgccgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5161 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5221 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5281 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5341 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5401 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5461 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5521 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5581 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5641 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5701 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5761 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5821 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5881 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5941 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6001 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6061 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6121 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6181 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6241 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6301 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctac gcgcgcgccg
     6361 gcggcgcgcc gtcgacggcc ggccgcggcc gc
//
