LOCUS       sequence-249232-        6483 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 6483)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6483)
  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-351 C-Tag Venus173/155[1] L20 {Esp3I(2x)...
FEATURES             Location/Qualifiers
     source          1..6483
                     /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(1641..2215,2216..2218,2219..2227)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1808..1825)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2035..2054
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2228..2230,2231..2233,2234..2947)
                     /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(2273..2294)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2993..3125
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3134..3722
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3187..3207)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3605..3616)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3749..3766)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3755..3866
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3962..3982)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3962..3980)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4001..4017)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4001..4017)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4014..4036)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4119..4138)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4184..4239
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4238..4257)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4618..5206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4698..4717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5377..6168,6169..6237))
                     /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     6000..6019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6238..6342)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6410..6428
                     /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 ccgtctccgg gagtcctggc ggtggatccg gaggctctgg
     1081 gagctctgcc agcggtggca gcaccagtgt gtcaaaggga gaagagttgt ttaccggtgt
     1141 tgttcccata ctggtggaac tggatggtga cgtcaatgga cacaagttta gcgtttctgg
     1201 cgaaggagaa ggcgatgcta cctatggcaa actgaccctc aaactgatct gtacgactgg
     1261 aaagttgcct gtaccttggc caacactggt cacaaccttg ggctatgggc tgcaatgctt
     1321 tgcccgttat cctgatcaca tgaagcaaca tgacttcttt aagtctgcta tgccagaggg
     1381 atatgtgcag gaacggacta tcttcttcaa ggacgacggc aactacaaga cgagagctga
     1441 ggtgaaattt gagggtgaca cactggtgaa tcgcatagag ctgaaaggca tcgacttcaa
     1501 agaagatggg aatattctcg ggcataaact cgagtacaac tacaattccc acaacgtgta
     1561 catcactgcc gataagcaga agaatgggat taaggcgaac ttcaagatcc gacacaacat
     1621 cgagtaagaa ttcagatccg cccctctccc tccccccccc ctaacgttac tggccgaagc
     1681 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     1741 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     1801 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     1861 ctggtagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     1921 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     1981 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     2041 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     2101 tctgatctgg ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     2161 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     2221 cacaaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2281 gctggacggc gacgtaaacg gccacaagtt cagcgtgagg ggcgagggcg agggcgatgc
     2341 caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2401 gcccaccctc gtgaccaccc tgagccacgg cgtgcagtgc ttcgcccgct accccgacca
     2461 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2521 catcttcttc aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga
     2581 caccctggtg aaccgcatcg agctgaaggg cgtcgacttc aaggaggacg gcaacatcct
     2641 ggggcacaag ctggagtaca acttcaacag ccacaacatc tatatcatgg ccgtcaagca
     2701 gaagaacggc atcaaggtga acttcaagat ccgccacaac gtggaggacg gcagcgtgca
     2761 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     2821 cagccactac ctgagcaccc agtccgtgct gagcaaagac cccaacgaga agcgcgatca
     2881 catggtcctg ctggagttcc gcaccgccgc cgggatcact ctcggcatgg acgagctgta
     2941 caagtaaacc ggtgctcgag gtaccgatga atagctaagg tcgaggccgc aggtaagtat
     3001 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
     3061 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
     3121 cacaggtgtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3181 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3241 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3301 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3361 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3421 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3481 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3541 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3601 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3661 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3721 gcctggaatt cgagctcggt acgatcagcc tcgactgtgc cttctagttg ccagccatct
     3781 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3841 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3901 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagccc agcttttgtt
     3961 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4021 gaaattgtta tccgctaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     4081 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     4141 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     4201 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     4261 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     4321 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     4381 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     4441 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     4501 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     4561 agatcggccg gccgccggcg tcgacgcgcg cggccgcggc gcgccgttgc tggcgttttt
     4621 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4681 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4741 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4801 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4861 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4921 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4981 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5041 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5101 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5161 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5221 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5281 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5341 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5401 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5461 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5521 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5581 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5641 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5701 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5761 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5821 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5881 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5941 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6001 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6061 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6121 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6181 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6241 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6301 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6361 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6421 cacgaggccc tttcgtctac gcgcgcgccg gcggcgcgcc gtcgacggcc ggccgcggcc
     6481 gcg
//
