LOCUS       sequence-249409-        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-552    N-Tag Venus210[2] L32 Tln1(rod) m...
FEATURES             Location/Qualifiers
     source          1..7142
                     /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"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(2310..2884,2885..2887,2888..2896)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2477..2494)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2704..2723
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2897..3607
                     /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(3042..3060)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3310..3329
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          3653..3785
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3794..4382
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3847..3867)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4265..4276)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4409..4426)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4415..4526
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4622..4642)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4622..4640)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4661..4677)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4661..4677)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4674..4696)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4779..4798)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4844..4899
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4898..4917)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5278..5866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5358..5377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6037..6828,6829..6897))
                     /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     6660..6679
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6898..7002)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7070..7088
                     /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 caccatggat cccaatgaga agagggacca tatggtactg ctcgaattcg tgactgcagc
     1081 aggcattacc cttgggatgg atgaacttta taaggacgtg ggtggcggag ggagcgaagg
     1141 cggcggatcc ggagggcctg gctctggcgg agaggggagc gctggaggcg ggtcagccgg
     1201 tggtgggagt gctgtgtctg gtatcattgc tgacctcgac accaccatca tgttcgctac
     1261 tgctggcaca cttaaccgtg agggtgctga aacttttgct gaccaccggg agggtatctt
     1321 aaagacagca aaggttcttg tggaggacac caaggtccta gtgcagaatg cagctgggag
     1381 ccaggagaag ttggcacaag ccgcccagtc ctccgtggcc accattaccc gcctcgctga
     1441 tgtggtcaag ctcggtgcag ccagcctagg agccgaagac cctgaaactc aggtggtgct
     1501 gatcaatgca gtaaaggacg tagccaaggc cctgggtgac ctcatcagcg ctacgaaggc
     1561 tgcagcgggc aaagttgggg atgaccctgc agtgtggcag ctcaagaact ctgccaaggt
     1621 gatggtgacc aatgtgacat cattgctcaa gacagtgaag gctgtggaag atgaggccac
     1681 caaaggcaca cgggccctag aggcaaccac agagcacata cgtcaggaac tggcggtctt
     1741 ctgttcccca gagccacctg ccaagacctc tacccctgaa gacttcatcc gaatgaccaa
     1801 gggtattact atggcaacag ccaaagccgt tgctgctggc aattcctgtc gacaggaaga
     1861 tgtcattgcc acagccaatc tgagccgacg ggctattgcg gacatgcttc gggcttgcaa
     1921 ggaagcagct ttccacccag aagtggcgcc tgatgtacgg ctccgagccc tgcactatgg
     1981 ccgggagtgt gccaatggtt acctggaact gctggaccac gtgctgctga cccttcagaa
     2041 gccaaaccca gacctgaagc agcagctaac tggacactca aagcgggttg ctggctctgt
     2101 gactgagctc atccaggctg ctgaagccat gaaaggaaca gagtgggtgg acccagagga
     2161 ccctactgtc attgctgaga atgagctctt gggagccgca gccgccatcg aggctgcagc
     2221 caagaagctg gagcagctga agcccagggc caaacccaag gaggcggatg agtccttgaa
     2281 ctttgaggaa caataagaat tcagatccgc ccctctccct cccccccccc taacgttact
     2341 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     2401 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     2461 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     2521 gcagttcctc tggtagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     2581 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     2641 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     2701 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     2761 tgtatgggat ctgatctggg gcctcggtac acatgcttta catgtgttta gtcgaggtta
     2821 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     2881 taatatggcc acaaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     2941 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     3001 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     3061 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     3121 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     3181 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     3241 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     3301 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     3361 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     3421 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     3481 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     3541 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     3601 caagtaaacc ggtgctcgag gtaccgatga atagctaagg tcgaggccgc aggtaagtat
     3661 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
     3721 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
     3781 cacaggtgtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3841 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3901 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3961 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4021 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4081 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4141 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     4201 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4261 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4321 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4381 gcctggaatt cgagctcggt acgatcagcc tcgactgtgc cttctagttg ccagccatct
     4441 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4501 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4561 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagccc agcttttgtt
     4621 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4681 gaaattgtta tccgctaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     4741 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     4801 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     4861 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     4921 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     4981 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     5041 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     5101 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     5161 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     5221 agatcggccg gccgccggcg tcgacgcgcg cggccgcggc gcgccgttgc tggcgttttt
     5281 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5341 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5401 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5461 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5521 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5581 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5641 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5701 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5761 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5821 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5881 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5941 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6001 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6061 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6121 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6181 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6241 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6301 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6361 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6421 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6481 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6541 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6601 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6661 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6721 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6781 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6841 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6901 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6961 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7021 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     7081 cacgaggccc tttcgtctac gcgcgcgccg gcggcgcgcc gtcgacggcc ggccgcggcc
     7141 gc
//
