LOCUS       sequence-249199-        6399 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 6399)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6399)
  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-318    N-Tag IFP1.4[1] L32 {Esp3I(2x)} E...
FEATURES             Location/Qualifiers
     source          1..6399
                     /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(1558..2132,2133..2135,2136..2144)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1725..1742)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1952..1971
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2145..2147,2148..2150,2151..2864)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(2190..2211)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2910..3042
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3051..3639
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3104..3124)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3522..3533)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3666..3683)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3672..3783
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3879..3899)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3879..3897)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3918..3934)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3918..3934)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3931..3953)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4036..4055)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4101..4156
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4155..4174)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4535..5123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4615..4634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5294..6085,6086..6154))
                     /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     5917..5936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6155..6259)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6327..6345
                     /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 caccatggct cgggatccat tgcctttctt cccacccttg tacctcggtg gaccagagat
     1081 caccaccgag aactgcgaga gggaaccgat acacattccc gggtccattc agccccatgg
     1141 tgcgctgctg actgctgacg gacattcagg ggaagtgctg caagtcagcc tgaatgcagc
     1201 aacctttctg ggccaagaac ccactgtgct tcgcgggcag accttggctg ctctgctgcc
     1261 tgagcagtgg ccagccctcc aagcagctct tccacccggt tgtccagacg ccctccagta
     1321 tcgtgccact ctggactggc ctgctgccgg ccaccttagc ctgacagtgc acagggtagc
     1381 tgagctgctc atcctcgaat tcgagcccac cgaggcatgg gacgtgggtg gcggagggag
     1441 cgaaggcggc ggatccggag ggcctggctc tggcggagag gggagcgctg gaggcgggtc
     1501 agccggtggt gggagtcgag acgccgaggc acgccgtctc ctaagaattc agatccgccc
     1561 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     1621 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     1681 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     1741 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gtagcttctt gaagacaaac
     1801 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     1861 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     1921 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     1981 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac
     2041 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     2101 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg
     2161 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2221 acaagttcag cgtgaggggc gagggcgagg gcgatgccac caacggcaag ctgaccctga
     2281 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2341 gccacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     2401 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2461 cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2521 tgaagggcgt cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2581 tcaacagcca caacatctat atcatggccg tcaagcagaa gaacggcatc aaggtgaact
     2641 tcaagatccg ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2701 acacccccat cggcgacggc cccgtgctgc tgcccgacag ccactacctg agcacccagt
     2761 ccgtgctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttccgca
     2821 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaaccggt gctcgaggta
     2881 ccgatgaata gctaaggtcg aggccgcagg taagtatcaa ggttacaaga caggtttaag
     2941 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     3001 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtcgac aatcaacctc
     3061 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3121 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3181 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3241 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3301 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3361 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3421 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     3481 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3541 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     3601 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacg
     3661 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3721 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3781 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3841 gggggaggat tgggaagaca atagcccagc ttttgttccc tttagtgagg gttaattgcg
     3901 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctaattcac
     3961 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     4021 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     4081 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     4141 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     4201 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     4261 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     4321 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     4381 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     4441 actactccca gtcatagctg tccctcttct cttatggaga tcggccggcc gccggcgtcg
     4501 acgcgcgcgg ccgcggcgcg ccgttgctgg cgtttttcca taggctccgc ccccctgacg
     4561 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4981 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5041 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5101 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5161 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5221 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5281 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5341 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5401 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5461 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5521 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5581 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5641 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5701 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5761 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5821 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5881 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5941 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6001 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6061 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6121 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6181 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6241 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6301 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctacgcg
     6361 cgcgccggcg gcgcgccgtc gacggccggc cgcggccgc
//
