LOCUS       sequence-249408-        7142 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 7142)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7142)
  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-551    C-Tag Venus210[2] L32 Tln1(rod) m...
FEATURES             Location/Qualifiers
     source          1..7142
                     /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(2309..2883,2884..2886,2887..2895)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2476..2493)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2703..2722
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2896..3606
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3041..3059)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3309..3328
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          3652..3784
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3793..4381
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3846..3866)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4264..4275)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4408..4425)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4414..4525
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4621..4641)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4621..4639)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4660..4676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4660..4676)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4673..4695)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4778..4797)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4843..4898
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4897..4916)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5277..5865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5357..5376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6036..6827,6828..6896))
                     /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     6659..6678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6897..7001)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7069..7087
                     /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 acgtgggtgg cggagggagc gaaggcggcg gatccggagg gcctggctct
     2161 ggcggagagg ggagcgctgg aggcgggtca gccggtggtg gcagtgatcc caatgagaag
     2221 agggaccata tggtactgct cgaattcgtg actgcagcag gcattaccct tgggatggat
     2281 gaactttata agtaagaatt cagatccgcc cctctccctc ccccccccct aacgttactg
     2341 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     2401 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     2461 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     2521 cagttcctct ggtagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     2581 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     2641 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     2701 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     2761 gtatgggatc tgatctgggg cctcggtaca catgctttac atgtgtttag tcgaggttaa
     2821 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     2881 aatatggcca caaccatggt gagcaagggc gaggaggata acatggccat catcaaggag
     2941 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     3001 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     3061 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     3121 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     3181 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     3241 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     3301 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     3361 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     3421 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     3481 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     3541 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     3601 aagtaaaccg gtgctcgagg taccgatgaa tagctaaggt cgaggccgca ggtaagtatc
     3661 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
     3721 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
     3781 acaggtgtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3841 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3901 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3961 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4021 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4081 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4141 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     4201 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4261 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4321 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4381 cctggaattc gagctcggta cgatcagcct cgactgtgcc ttctagttgc cagccatctg
     4441 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     4501 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     4561 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagccca gcttttgttc
     4621 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4681 aaattgttat ccgctaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg
     4741 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     4801 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     4861 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     4921 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     4981 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     5041 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     5101 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     5161 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     5221 gatcggccgg ccgccggcgt cgacgcgcgc ggccgcggcg cgccgttgct ggcgtttttc
     5281 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5341 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5401 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5461 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5521 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5581 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5641 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5701 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5761 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5821 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5881 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5941 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6001 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6061 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6121 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6181 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6241 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6301 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6361 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6421 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6481 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6541 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6601 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6661 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6721 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6781 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6841 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6901 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6961 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7021 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     7081 acgaggccct ttcgtctacg cgcgcgccgg cggcgcgccg tcgacggccg gccgcggccg
     7141 cg
//
