LOCUS       sequence-270544-        7655 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 7655)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7655)
  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-543    C-Tag Venus210[1] L20 Tln1(rod) E...
FEATURES             Location/Qualifiers
     source          1..7655
                     /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"
     regulatory      1018..1027
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(2813..3387,3388..3390,3391..3399)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2980..2997)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3207..3226
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3400..3402,3403..3405,3406..4119)
                     /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(3445..3466)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          4165..4297
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    4306..4894
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4359..4379)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4777..4788)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4921..4938)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4927..5038
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5134..5154)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5134..5152)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5173..5189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5173..5189)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5186..5208)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(5291..5310)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5356..5411
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5410..5429)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5790..6378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5870..5889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6549..7340,7341..7409))
                     /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     7172..7191
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7410..7514)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7582..7600
                     /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 accatggctg tgtctggtat cattgctgac ctcgacacca ccatcatgtt cgctactgct
     1081 ggcacactta accgtgaggg tgctgaaact tttgctgacc accgggaggg tatcttaaag
     1141 acagcaaagg ttcttgtgga ggacaccaag gtcctagtgc agaatgcagc tgggagccag
     1201 gagaagttgg cacaagccgc ccagtcctcc gtggccacca ttacccgcct cgctgatgtg
     1261 gtcaagctcg gtgcagccag cctaggagcc gaagaccctg aaactcaggt ggtgctgatc
     1321 aatgcagtaa aggacgtagc caaggccctg ggtgacctca tcagcgctac gaaggctgca
     1381 gcgggcaaag ttggggatga ccctgcagtg tggcagctca agaactctgc caaggtgatg
     1441 gtgaccaatg tgacatcatt gctcaagaca gtgaaggctg tggaagatga ggccaccaaa
     1501 ggcacacggg ccctagaggc aaccacagag cacatacgtc aggaactggc ggtcttctgt
     1561 tccccagagc cacctgccaa gacctctacc cctgaagact tcatccgaat gaccaagggt
     1621 attactatgg caacagccaa agccgttgct gctggcaatt cctgtcgaca ggaagatgtc
     1681 attgccacag ccaatctgag ccgacgggct attgcggaca tgcttcgggc ttgcaaggaa
     1741 gcagctttcc acccagaagt ggcgcctgat gtacggctcc gagccctgca ctatggccgg
     1801 gagtgtgcca atggttacct ggaactgctg gaccacgtgc tgctgaccct tcagaagcca
     1861 aacccagacc tgaagcagca gctaactgga cactcaaagc gggttgctgg ctctgtgact
     1921 gagctcatcc aggctgctga agccatgaaa ggaacagagt gggtggaccc agaggaccct
     1981 actgtcattg ctgagaatga gctcttggga gccgcagccg ccatcgaggc tgcagccaag
     2041 aagctggagc agctgaagcc cagggccaaa cccaaggagg cggatgagtc cttgaacttt
     2101 gaggaacaag ggagtcctgg cggtggatcc ggaggctctg ggagctctgc cagcggtggc
     2161 agcaccagtg tgtcaaaggg agaagagttg tttaccggtg ttgttcccat actggtggaa
     2221 ctggatggtg acgtcaatgg acacaagttt agcgtttctg gcgaaggaga aggcgatgct
     2281 acctatggca aactgaccct caaactgatc tgtacgactg gaaagttgcc tgtaccttgg
     2341 ccaacactgg tcacaacctt gggctatggg ctgcaatgct ttgcccgtta tcctgatcac
     2401 atgaagcaac atgacttctt taagtctgct atgccagagg gatatgtgca ggaacggact
     2461 atcttcttca aggacgacgg caactacaag acgagagctg aggtgaaatt tgagggtgac
     2521 acactggtga atcgcataga gctgaaaggc atcgacttca aagaagatgg gaatattctc
     2581 gggcataaac tcgagtacaa ctacaattcc cacaacgtgt acatcactgc cgataagcag
     2641 aagaatggga ttaaggcgaa cttcaagatc cgacacaaca tcgaggatgg tggggtccag
     2701 cttgccgacc attatcagca gaacacaccc attggggatg gaccagtcct gttgccggat
     2761 aaccactacc tcagttacca gagcaaactg tccaaataag aattcagatc cgcccctctc
     2821 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     2881 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     2941 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     3001 gtctgttgaa tgtcgtgaag gaagcagttc ctctggtagc ttcttgaaga caaacaacgt
     3061 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     3121 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     3181 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     3241 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     3301 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     3361 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     3421 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3481 ttcagcgtga ggggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc
     3541 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgagccac
     3601 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3661 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcacctac
     3721 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3781 ggcgtcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac
     3841 agccacaaca tctatatcat ggccgtcaag cagaagaacg gcatcaaggt gaacttcaag
     3901 atccgccaca acgtggagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     3961 cccatcggcg acggccccgt gctgctgccc gacagccact acctgagcac ccagtccgtg
     4021 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt ccgcaccgcc
     4081 gccgggatca ctctcggcat ggacgagctg tacaagtaaa ccggtgctcg aggtaccgat
     4141 gaatagctaa ggtcgaggcc gcaggtaagt atcaaggtta caagacaggt ttaaggagac
     4201 caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt
     4261 ggtcttactg acatccactt tgcctttctc tccacaggtg tcgacaatca acctctggat
     4321 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     4381 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     4441 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     4501 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     4561 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     4621 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     4681 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     4741 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     4801 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     4861 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg gtacgatcag
     4921 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4981 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     5041 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     5101 aggattggga agacaatagc ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     5161 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctaa ttcactcctc
     5221 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     5281 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     5341 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     5401 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     5461 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     5521 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     5581 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     5641 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     5701 tcccagtcat agctgtccct cttctcttat ggagatcggc cggccgccgg cgtcgacgcg
     5761 cgcggccgcg gcgcgccgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5821 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5881 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5941 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6001 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6061 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6121 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6181 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6241 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6301 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6361 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6421 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6481 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6541 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6601 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6661 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6721 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6781 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6841 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6901 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6961 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7021 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7081 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7141 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7201 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7261 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7321 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7381 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7441 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7501 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     7561 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct acgcgcgcgc
     7621 cggcggcgcg ccgtcgacgg ccggccgcgg ccgcg
//
