LOCUS       sequence-249242-        6564 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 6564)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6564)
  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-361    C-Tag Venus210[1] L10 {Esp3I(2x)}...
FEATURES             Location/Qualifiers
     source          1..6564
                     /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(1723..2297,2298..2300,2301..2309)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1890..1907)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2117..2136
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2310..2312,2313..2315,2316..3029)
                     /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(2355..2376)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3075..3207
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3216..3804
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3269..3289)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3687..3698)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3831..3848)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3837..3948
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4044..4064)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4044..4062)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4083..4099)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4083..4099)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4096..4118)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4201..4220)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4266..4321
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4320..4339)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4700..5288)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4780..4799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5459..6250,6251..6319))
                     /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     6082..6101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6320..6424)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6492..6510
                     /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 tgtcaaaggg agaagagttg tttaccggtg ttgttcccat actggtggaa ctggatggtg
     1141 acgtcaatgg acacaagttt agcgtttctg gcgaaggaga aggcgatgct acctatggca
     1201 aactgaccct caaactgatc tgtacgactg gaaagttgcc tgtaccttgg ccaacactgg
     1261 tcacaacctt gggctatggg ctgcaatgct ttgcccgtta tcctgatcac atgaagcaac
     1321 atgacttctt taagtctgct atgccagagg gatatgtgca ggaacggact atcttcttca
     1381 aggacgacgg caactacaag acgagagctg aggtgaaatt tgagggtgac acactggtga
     1441 atcgcataga gctgaaaggc atcgacttca aagaagatgg gaatattctc gggcataaac
     1501 tcgagtacaa ctacaattcc cacaacgtgt acatcactgc cgataagcag aagaatggga
     1561 ttaaggcgaa cttcaagatc cgacacaaca tcgaggatgg tggggtccag cttgccgacc
     1621 attatcagca gaacacaccc attggggatg gaccagtcct gttgccggat aaccactacc
     1681 tcagttacca gagcaaactg tccaaataag aattcagatc cgcccctctc cctccccccc
     1741 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     1801 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     1861 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     1921 tgtcgtgaag gaagcagttc ctctggtagc ttcttgaaga caaacaacgt ctgtagcgac
     1981 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     2041 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     2101 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     2161 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt
     2221 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     2281 aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag ctgttcaccg
     2341 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtga
     2401 ggggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca
     2461 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgagccac ggcgtgcagt
     2521 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2581 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcacctac aagacccgcg
     2641 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcgtcgact
     2701 tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac agccacaaca
     2761 tctatatcat ggccgtcaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     2821 acgtggagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2881 acggccccgt gctgctgccc gacagccact acctgagcac ccagtccgtg ctgagcaaag
     2941 accccaacga gaagcgcgat cacatggtcc tgctggagtt ccgcaccgcc gccgggatca
     3001 ctctcggcat ggacgagctg tacaagtaaa ccggtgctcg aggtaccgat gaatagctaa
     3061 ggtcgaggcc gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac
     3121 tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg
     3181 acatccactt tgcctttctc tccacaggtg tcgacaatca acctctggat tacaaaattt
     3241 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3301 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3361 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3421 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3481 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3541 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3601 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     3661 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3721 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3781 tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg gtacgatcag cctcgactgt
     3841 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3901 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3961 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4021 agacaatagc ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     4081 tggtcatagc tgtttcctgt gtgaaattgt tatccgctaa ttcactcctc aggtgcaggc
     4141 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     4201 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     4261 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     4321 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     4381 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     4441 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     4501 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     4561 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     4621 agctgtccct cttctcttat ggagatcggc cggccgccgg cgtcgacgcg cgcggccgcg
     4681 gcgcgccgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4741 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4801 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4861 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4921 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4981 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5041 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5101 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5161 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5221 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5281 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5341 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5401 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5461 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5521 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5581 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5641 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5701 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5761 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5821 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5881 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5941 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6001 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6061 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6121 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6181 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6241 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6301 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6361 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6421 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     6481 taacctataa aaataggcgt atcacgaggc cctttcgtct acgcgcgcgc cggcggcgcg
     6541 ccgtcgacgg ccggccgcgg ccgc
//
