LOCUS       sequence-249403-        7691 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 7691)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7691)
  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-546    N-Tag Venus210[1] L32 Tln1(rod) E...
FEATURES             Location/Qualifiers
     source          1..7691
                     /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(2850..3424,3425..3427,3428..3436)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3017..3034)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3244..3263
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3437..3439,3440..3442,3443..4156)
                     /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(3482..3503)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          4202..4334
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    4343..4931
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4396..4416)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4814..4825)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4958..4975)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4964..5075
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5171..5191)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5171..5189)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5210..5226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5210..5226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5223..5245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(5328..5347)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5393..5448
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5447..5466)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5827..6415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5907..5926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6586..7377,7378..7446))
                     /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     7209..7228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7447..7551)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7619..7637
                     /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 caccatggtg tcaaagggag aagagttgtt taccggtgtt gttcccatac tggtggaact
     1081 ggatggtgac gtcaatggac acaagtttag cgtttctggc gaaggagaag gcgatgctac
     1141 ctatggcaaa ctgaccctca aactgatctg tacgactgga aagttgcctg taccttggcc
     1201 aacactggtc acaaccttgg gctatgggct gcaatgcttt gcccgttatc ctgatcacat
     1261 gaagcaacat gacttcttta agtctgctat gccagaggga tatgtgcagg aacggactat
     1321 cttcttcaag gacgacggca actacaagac gagagctgag gtgaaatttg agggtgacac
     1381 actggtgaat cgcatagagc tgaaaggcat cgacttcaaa gaagatggga atattctcgg
     1441 gcataaactc gagtacaact acaattccca caacgtgtac atcactgccg ataagcagaa
     1501 gaatgggatt aaggcgaact tcaagatccg acacaacatc gaggatggtg gggtccagct
     1561 tgccgaccat tatcagcaga acacacccat tggggatgga ccagtcctgt tgccggataa
     1621 ccactacctc agttaccaga gcaaactgtc caaagacgtg ggtggcggag ggagcgaagg
     1681 cggcggatcc ggagggcctg gctctggcgg agaggggagc gctggaggcg ggtcagccgg
     1741 tggtgggagt gctgtgtctg gtatcattgc tgacctcgac accaccatca tgttcgctac
     1801 tgctggcaca cttaaccgtg agggtgctga aacttttgct gaccaccggg agggtatctt
     1861 aaagacagca aaggttcttg tggaggacac caaggtccta gtgcagaatg cagctgggag
     1921 ccaggagaag ttggcacaag ccgcccagtc ctccgtggcc accattaccc gcctcgctga
     1981 tgtggtcaag ctcggtgcag ccagcctagg agccgaagac cctgaaactc aggtggtgct
     2041 gatcaatgca gtaaaggacg tagccaaggc cctgggtgac ctcatcagcg ctacgaaggc
     2101 tgcagcgggc aaagttgggg atgaccctgc agtgtggcag ctcaagaact ctgccaaggt
     2161 gatggtgacc aatgtgacat cattgctcaa gacagtgaag gctgtggaag atgaggccac
     2221 caaaggcaca cgggccctag aggcaaccac agagcacata cgtcaggaac tggcggtctt
     2281 ctgttcccca gagccacctg ccaagacctc tacccctgaa gacttcatcc gaatgaccaa
     2341 gggtattact atggcaacag ccaaagccgt tgctgctggc aattcctgtc gacaggaaga
     2401 tgtcattgcc acagccaatc tgagccgacg ggctattgcg gacatgcttc gggcttgcaa
     2461 ggaagcagct ttccacccag aagtggcgcc tgatgtacgg ctccgagccc tgcactatgg
     2521 ccgggagtgt gccaatggtt acctggaact gctggaccac gtgctgctga cccttcagaa
     2581 gccaaaccca gacctgaagc agcagctaac tggacactca aagcgggttg ctggctctgt
     2641 gactgagctc atccaggctg ctgaagccat gaaaggaaca gagtgggtgg acccagagga
     2701 ccctactgtc attgctgaga atgagctctt gggagccgca gccgccatcg aggctgcagc
     2761 caagaagctg gagcagctga agcccagggc caaacccaag gaggcggatg agtccttgaa
     2821 ctttgaggaa caataagaat tcagatccgc ccctctccct cccccccccc taacgttact
     2881 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     2941 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     3001 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     3061 gcagttcctc tggtagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     3121 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     3181 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     3241 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     3301 tgtatgggat ctgatctggg gcctcggtac acatgcttta catgtgttta gtcgaggtta
     3361 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     3421 taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3481 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgaggg gcgagggcga
     3541 gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3601 cgtgccctgg cccaccctcg tgaccaccct gagccacggc gtgcagtgct tcgcccgcta
     3661 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3721 ggagcgcacc atcttcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt
     3781 cgagggcgac accctggtga accgcatcga gctgaagggc gtcgacttca aggaggacgg
     3841 caacatcctg gggcacaagc tggagtacaa cttcaacagc cacaacatct atatcatggc
     3901 cgtcaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaacg tggaggacgg
     3961 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4021 gctgcccgac agccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa
     4081 gcgcgatcac atggtcctgc tggagttccg caccgccgcc gggatcactc tcggcatgga
     4141 cgagctgtac aagtaaaccg gtgctcgagg taccgatgaa tagctaaggt cgaggccgca
     4201 ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag
     4261 acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc
     4321 ctttctctcc acaggtgtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4381 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4441 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4501 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4561 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4621 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4681 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     4741 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4801 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4861 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4921 cgcctccccg cctggaattc gagctcggta cgatcagcct cgactgtgcc ttctagttgc
     4981 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     5041 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     5101 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagccca
     5161 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     5221 ttcctgtgtg aaattgttat ccgctaattc actcctcagg tgcaggctgc ctatcagaag
     5281 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     5341 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     5401 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     5461 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     5521 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     5581 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     5641 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     5701 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     5761 ctcttatgga gatcggccgg ccgccggcgt cgacgcgcgc ggccgcggcg cgccgttgct
     5821 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5881 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5941 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6001 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6061 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6121 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6181 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6241 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6301 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6361 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6421 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6481 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6541 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6601 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6661 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6721 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6781 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6841 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6901 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6961 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7021 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7081 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7141 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7201 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7261 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7321 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7381 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7441 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7501 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7561 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     7621 taggcgtatc acgaggccct ttcgtctacg cgcgcgccgg cggcgcgccg tcgacggccg
     7681 gccgcggccg c
//
