LOCUS       sequence-249433-        7112 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 7112)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7112)
  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-528    N-Tag GFP[1] L32 Tln1(rod) mCherry
FEATURES             Location/Qualifiers
     source          1..7112
                     /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"
     misc_feature    join(2280..2854,2855..2857,2858..2866)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2447..2464)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2674..2693
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2867..3577
                     /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(3012..3030)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3280..3299
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          3623..3755
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3764..4352
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3817..3837)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4235..4246)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4379..4396)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4385..4496
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4592..4612)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4592..4610)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4631..4647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4631..4647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4644..4666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4749..4768)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4814..4869
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4868..4887)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5248..5836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5328..5347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6007..6798,6799..6867))
                     /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     6630..6649
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6868..6972)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7040..7058
                     /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 ctgcctgatg accactacct gagcacacag accatcctct ccaaggacct
     1081 taacgacgtg ggtggcggag ggagcgaagg cggcggatcc ggagggcctg gctctggcgg
     1141 agaggggagc gctggaggcg ggtcagccgg tggtgggagt gctgtgtctg gtatcattgc
     1201 tgacctcgac accaccatca tgttcgctac tgctggcaca cttaaccgtg agggtgctga
     1261 aacttttgct gaccaccggg agggtatctt aaagacagca aaggttcttg tggaggacac
     1321 caaggtccta gtgcagaatg cagctgggag ccaggagaag ttggcacaag ccgcccagtc
     1381 ctccgtggcc accattaccc gcctcgctga tgtggtcaag ctcggtgcag ccagcctagg
     1441 agccgaagac cctgaaactc aggtggtgct gatcaatgca gtaaaggacg tagccaaggc
     1501 cctgggtgac ctcatcagcg ctacgaaggc tgcagcgggc aaagttgggg atgaccctgc
     1561 agtgtggcag ctcaagaact ctgccaaggt gatggtgacc aatgtgacat cattgctcaa
     1621 gacagtgaag gctgtggaag atgaggccac caaaggcaca cgggccctag aggcaaccac
     1681 agagcacata cgtcaggaac tggcggtctt ctgttcccca gagccacctg ccaagacctc
     1741 tacccctgaa gacttcatcc gaatgaccaa gggtattact atggcaacag ccaaagccgt
     1801 tgctgctggc aattcctgtc gacaggaaga tgtcattgcc acagccaatc tgagccgacg
     1861 ggctattgcg gacatgcttc gggcttgcaa ggaagcagct ttccacccag aagtggcgcc
     1921 tgatgtacgg ctccgagccc tgcactatgg ccgggagtgt gccaatggtt acctggaact
     1981 gctggaccac gtgctgctga cccttcagaa gccaaaccca gacctgaagc agcagctaac
     2041 tggacactca aagcgggttg ctggctctgt gactgagctc atccaggctg ctgaagccat
     2101 gaaaggaaca gagtgggtgg acccagagga ccctactgtc attgctgaga atgagctctt
     2161 gggagccgca gccgccatcg aggctgcagc caagaagctg gagcagctga agcccagggc
     2221 caaacccaag gaggcggatg agtccttgaa ctttgaggaa caataagaat tcagatccgc
     2281 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     2341 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     2401 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     2461 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggtagcttc ttgaagacaa
     2521 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     2581 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     2641 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     2701 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac
     2761 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     2821 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg
     2881 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     2941 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     3001 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     3061 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     3121 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     3181 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     3241 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     3301 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     3361 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     3421 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     3481 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     3541 ccactccacc ggcggcatgg acgagctgta caagtaaacc ggtgctcgag gtaccgatga
     3601 atagctaagg tcgaggccgc aggtaagtat caaggttaca agacaggttt aaggagacca
     3661 atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg
     3721 tcttactgac atccactttg cctttctctc cacaggtgtc gacaatcaac ctctggatta
     3781 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3841 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3901 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3961 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4021 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4081 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4141 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     4201 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4261 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4321 gagtcggatc tccctttggg ccgcctcccc gcctggaatt cgagctcggt acgatcagcc
     4381 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4441 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4501 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4561 gattgggaag acaatagccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     4621 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctaatt cactcctcag
     4681 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     4741 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     4801 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     4861 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     4921 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     4981 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     5041 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     5101 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     5161 ccagtcatag ctgtccctct tctcttatgg agatcggccg gccgccggcg tcgacgcgcg
     5221 cggccgcggc gcgccgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5941 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6001 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6061 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6121 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6181 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6241 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6301 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6361 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6421 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6481 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6541 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6601 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6661 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6721 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6781 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6841 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6901 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6961 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     7021 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctac gcgcgcgccg
     7081 gcggcgcgcc gtcgacggcc ggccgcggcc gc
//
