LOCUS       sequence-265030-        6519 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 6519)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6519)
  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-353 C-Tag Venus173/155[1] L32 {Esp3I(2x)...
FEATURES             Location/Qualifiers
     source          1..6519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     732..756
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          871..1003
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    join(1677..2251,2252..2254,2255..2263)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1844..1861)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2071..2090
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2264..2266,2267..2269,2270..2983)
                     /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(2309..2330)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3029..3161
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3170..3758
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3223..3243)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3641..3652)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3785..3802)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3791..3902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3998..4018)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3998..4016)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4037..4053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4037..4053)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4050..4072)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4155..4174)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4220..4275
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4274..4293)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4654..5242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4734..4753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5413..6204,6205..6273))
                     /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     6036..6055
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6274..6378)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6446..6464
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctgcctcca ggcctcaata ttggccatta gccatattat tcattggtta tatagcataa
       61 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
      121 attggctcat gtccaatatg accgccatgt tggcattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt
      781 tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc
      841 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
      901 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
      961 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccaggcc
     1021 acccgagacg ccgaggcacg ccgtctccga cgtgggtggc ggagggagcg aaggcggcgg
     1081 atccggaggg cctggctctg gcggagaggg gagcgctgga ggcgggtcag ccggtggtgg
     1141 cagtgtgtca aagggagaag agttgtttac cggtgttgtt cccatactgg tggaactgga
     1201 tggtgacgtc aatggacaca agtttagcgt ttctggcgaa ggagaaggcg atgctaccta
     1261 tggcaaactg accctcaaac tgatctgtac gactggaaag ttgcctgtac cttggccaac
     1321 actggtcaca accttgggct atgggctgca atgctttgcc cgttatcctg atcacatgaa
     1381 gcaacatgac ttctttaagt ctgctatgcc agagggatat gtgcaggaac ggactatctt
     1441 cttcaaggac gacggcaact acaagacgag agctgaggtg aaatttgagg gtgacacact
     1501 ggtgaatcgc atagagctga aaggcatcga cttcaaagaa gatgggaata ttctcgggca
     1561 taaactcgag tacaactaca attcccacaa cgtgtacatc actgccgata agcagaagaa
     1621 tgggattaag gcgaacttca agatccgaca caacatcgag taagaattca gatccgcccc
     1681 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     1741 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     1801 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     1861 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg tagcttcttg aagacaaaca
     1921 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     1981 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     2041 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     2101 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtacaca
     2161 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     2221 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga
     2281 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2341 caagttcagc gtgaggggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa
     2401 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgag
     2461 ccacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2521 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcac
     2581 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2641 gaagggcgtc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt
     2701 caacagccac aacatctata tcatggccgt caagcagaag aacggcatca aggtgaactt
     2761 caagatccgc cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2821 cacccccatc ggcgacggcc ccgtgctgct gcccgacagc cactacctga gcacccagtc
     2881 cgtgctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttccgcac
     2941 cgccgccggg atcactctcg gcatggacga gctgtacaag taaaccggtg ctcgaggtac
     3001 cgatgaatag ctaaggtcga ggccgcaggt aagtatcaag gttacaagac aggtttaagg
     3061 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     3121 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtcgaca atcaacctct
     3181 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3241 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3301 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3361 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3421 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3481 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3541 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3601 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3661 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3721 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacga
     3781 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3841 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3901 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3961 ggggaggatt gggaagacaa tagcccagct tttgttccct ttagtgaggg ttaattgcgc
     4021 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact
     4081 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     4141 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     4201 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     4261 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     4321 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     4381 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     4441 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     4501 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     4561 ctactcccag tcatagctgt ccctcttctc ttatggagat cggccggccg ccggcgtcga
     4621 cgcgcgcggc cgcggcgcgc cgttgctggc gtttttccat aggctccgcc cccctgacga
     4681 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5041 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5101 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5161 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5221 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5281 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5341 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5401 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5461 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5521 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5581 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5641 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5701 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5761 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5821 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5881 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5941 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6001 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6061 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6121 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6181 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6241 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6301 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6361 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6421 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctacgcgc
     6481 gcgccggcgg cgcgccgtcg acggccggcc gcggccgcg
//
