LOCUS       sequence-249289-        6884 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 6884)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6884)
  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-122    N-Tag miRFP670[2] L32 FKBP EGFP
FEATURES             Location/Qualifiers
     source          1..6884
                     /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             1706..2029
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(2043..2617,2618..2620,2621..2629)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2210..2227)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2437..2456
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2630..2632,2633..2635,2636..3349)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2630..2632,2633..2635,2636..3346)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2675..2696)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2936..2955)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3283..3304
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3395..3527
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3536..4124
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3589..3609)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4007..4018)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4151..4168)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4157..4268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4364..4384)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4364..4382)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4403..4419)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4403..4419)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4416..4438)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4521..4540)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4586..4641
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4640..4659)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5020..5608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5100..5119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5779..6570,6571..6639))
                     /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     6402..6421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6640..6744)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6812..6830
                     /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 caccatgggc agagcgggac ccagtatcga cctgtctggg acactcgcac cagcactgga
     1081 gcgcataaga actgctggat cactgagggc cctctgtgac gacacagtct tgctctttca
     1141 gcaatgtacg ggctatgatc gagtgatggt gtaccgattc gacgaacagg gtcatggtct
     1201 ggtcttttcc gagtgtcacg tgcctggcct ggagagctac tttggcaacc gctatccctc
     1261 ttctaccgta ccccagatgg ccaggcaact ttacgtccgg caacgggtca gagtgctggt
     1321 ggatgtgacc taccagcctg tgccactcga acctcggctg agcccactca ctggacggga
     1381 tctggacatg agcggctgct tcctgaggtc catgtcacct tgccatctgc agttcttgaa
     1441 ggacatgggc gttcgagcca cactcgcagt ctcactggtt gtaggcggga aactgtgggg
     1501 tcttgtggtg tgtcaccact accttcccag gttcatccgg ttcgagttga gagccatctg
     1561 caaacgtttg gccgagagaa tagccacccg cattactgct ctggaatccg acgtgggtgg
     1621 cggagggagc gaaggcggcg gatccggagg gcctggctct ggcggagagg ggagcgctgg
     1681 aggcgggtca gccggtggtg ggagtggagt gcaggtggaa accatctccc caggagacgg
     1741 gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac tacaccggga tgcttgaaga
     1801 tggaaagaaa tttgattcct cccgggacag aaacaagccc tttaagttta tgctaggcaa
     1861 gcaggaggtg atccgaggct gggaagaagg ggttgcccag atgagtgtgg gtcagagagc
     1921 caaactgact atatctccag attatgccta tggtgccact gggcacccag gcatcatccc
     1981 accacatgcc actctcgtct tcgatgtgga gcttctaaaa ctggaataag aattcagatc
     2041 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     2101 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     2161 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     2221 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggtagc ttcttgaaga
     2281 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     2341 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     2401 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     2461 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     2521 tacacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     2581 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa
     2641 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2701 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2761 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2821 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2881 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2941 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3001 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3061 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     3121 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3181 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     3241 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3301 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa ccggtgctcg
     3361 aggtaccgat gaatagctaa ggtcgaggcc gcaggtaagt atcaaggtta caagacaggt
     3421 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     3481 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tcgacaatca
     3541 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3601 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3661 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3721 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3781 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3841 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3901 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3961 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4021 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4081 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg
     4141 gtacgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4201 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4261 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4321 agcaaggggg aggattggga agacaatagc ccagcttttg ttccctttag tgagggttaa
     4381 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctaa
     4441 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     4501 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     4561 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     4621 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     4681 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     4741 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     4801 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     4861 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     4921 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatcggc cggccgccgg
     4981 cgtcgacgcg cgcggccgcg gcgcgccgtt gctggcgttt ttccataggc tccgcccccc
     5041 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5101 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5161 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5221 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5281 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5341 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5401 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5461 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5521 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5581 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5641 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5701 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5761 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5821 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5881 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5941 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6001 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6061 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6121 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6181 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6241 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6301 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6361 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6421 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6481 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6541 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6601 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6661 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6721 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6781 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6841 acgcgcgcgc cggcggcgcg ccgtcgacgg ccggccgcgg ccgc
//
