LOCUS       sequence-249200-        6504 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 6504)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6504)
  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-319    C-Tag IFP1.4[2] L10 {Esp3I(2x)} EGFP
FEATURES             Location/Qualifiers
     source          1..6504
                     /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"
     misc_feature    join(1663..2237,2238..2240,2241..2249)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1830..1847)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2057..2076
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2250..2252,2253..2255,2256..2969)
                     /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(2250..2252,2253..2255,2256..2966)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2295..2316)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2556..2575)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2903..2924
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3015..3147
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3156..3744
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3209..3229)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3627..3638)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3771..3788)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3777..3888
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3984..4004)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3984..4002)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4023..4039)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4023..4039)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4036..4058)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4141..4160)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4206..4261
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4260..4279)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4640..5228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4720..4739)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5399..6190,6191..6259))
                     /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     6022..6041
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6260..6364)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6432..6450
                     /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 cacccgagac gccgaggcac gccgtctccg ggagctctgc cagcggtggc agcaccagtg
     1081 acagcattgg accccatgcg ctgaggaacg ctatgttcgc ccttgaatct gctcccaatc
     1141 tccgagcact ggctgaagtc gccacacaga cagtcaggga gctgacaggc tttgatcggg
     1201 tgatgctgta caagtttgcc ccagatgcaa cgggtgagat gatcgcagaa gccagacggg
     1261 aagggatgca ggccttcctg ggccacagat ttccggcctc acatacgcct gctcaggcca
     1321 gagcactgta cacccgacac ctcctgagac tgactgcgga cactagagca gctgccgtgc
     1381 cactcgatcc cgtgctcaac ccacagacca atgcgcctac ccctttggga ggagccgtac
     1441 tgagggccac atcccctatg cacatgcagt atctgcgcaa catgggcgtt gggagtagtc
     1501 tgtctgtgtc cgttgtcgtt ggagggcaac tgtggggctt gatcgtctgc caccatcaga
     1561 caccgtatgt gcttcctcct gacttgcgta caactctgga ggagctgggc cgcaagctta
     1621 gcggccaggt gcagcggaaa gaggcctaag aattcagatc cgcccctctc cctccccccc
     1681 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     1741 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     1801 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     1861 tgtcgtgaag gaagcagttc ctctggtagc ttcttgaaga caaacaacgt ctgtagcgac
     1921 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     1981 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     2041 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     2101 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt
     2161 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     2221 aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag ctgttcaccg
     2281 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2341 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2401 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2461 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2521 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2581 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2641 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     2701 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     2761 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2821 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     2881 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2941 ctctcggcat ggacgagctg tacaagtaaa ccggtgctcg aggtaccgat gaatagctaa
     3001 ggtcgaggcc gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac
     3061 tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg
     3121 acatccactt tgcctttctc tccacaggtg tcgacaatca acctctggat tacaaaattt
     3181 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3241 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3301 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3361 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3421 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3481 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3541 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     3601 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3661 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3721 tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg gtacgatcag cctcgactgt
     3781 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3841 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3901 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3961 agacaatagc ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     4021 tggtcatagc tgtttcctgt gtgaaattgt tatccgctaa ttcactcctc aggtgcaggc
     4081 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     4141 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     4201 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     4261 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     4321 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     4381 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     4441 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     4501 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     4561 agctgtccct cttctcttat ggagatcggc cggccgccgg cgtcgacgcg cgcggccgcg
     4621 gcgcgccgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4681 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4741 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4801 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4861 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4921 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4981 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5041 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5101 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5161 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5221 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5281 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5341 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5401 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5461 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5521 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5581 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5641 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5701 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5761 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5821 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5881 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5941 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6001 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6061 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6121 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6181 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6241 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6301 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6361 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     6421 taacctataa aaataggcgt atcacgaggc cctttcgtct acgcgcgcgc cggcggcgcg
     6481 ccgtcgacgg ccggccgcgg ccgc
//
