LOCUS       sequence-271319-        8879 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 8879)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8879)
  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-616 N-Tag Venus210[1] L20 Stat3(R609Q) E...
FEATURES             Location/Qualifiers
     source          1..8879
                     /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(4038..4612,4613..4615,4616..4624)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4205..4222)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4432..4451
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4625..4627,4628..4630,4631..5344)
                     /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(4670..4691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          5390..5522
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5531..6119
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5584..5604)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6002..6013)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6146..6163)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6152..6263
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6359..6379)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6359..6377)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6398..6414)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6398..6414)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6411..6433)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6516..6535)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6581..6636
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6635..6654)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(7015..7603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7095..7114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7774..8565,8566..8634))
                     /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     8397..8416
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8635..8739)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8807..8825
                     /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 caaagggagt cctggcggtg gatccggagg
     1681 ctctgggagc tctgccagcg gtggcagcac ctccgctcag tggaaccagc tgcagcagct
     1741 ggacacacgc tacctggagc agctgcacca gctgtacagc gacagcttcc ccatggagct
     1801 gcggcagttc ctggcacctt ggattgagag tcaagactgg gcatatgcag ccagcaaaga
     1861 gtcacatgcc acgttggtgt ttcataatct cttgggtgaa attgaccagc aatatagccg
     1921 attcctgcaa gagtccaatg tcctctatca gcacaacctt cgaagaatca agcagtttct
     1981 gcagagcagg tatcttgaga agccaatgga aattgcccgg attgtggccc gatgcctgtg
     2041 ggaagagtct cgcctcctcc agacggcagc cacggcagcc cagcaagggg gccaggccaa
     2101 ccacccaaca gccgccgtag tgacagagaa gcagcagatg ttggagcagc atcttcagga
     2161 tgtccggaag cgagtgcagg atctagaaca gaaaatgaag gtggtggaga acctccagga
     2221 cgactttgat ttcaactaca aaaccctcaa gagccaagga gacatgcagg atctgaatgg
     2281 aaacaaccag tctgtgacca gacagaagat gcagcagctg gaacagatgc tcacagccct
     2341 ggaccagatg cggagaagca ttgtgagtga gctggcgggg ctcttgtcag caatggagta
     2401 cgtgcagaag acactgactg atgaagagct ggctgactgg aagaggcggc agcagatcgc
     2461 gtgcatcgga ggccctccca acatctgcct ggaccgtctg gaaaactgga taacttcatt
     2521 agcagaatct caacttcaga cccgccaaca aattaagaaa ctggaggagc tgcagcagaa
     2581 agtgtcctac aagggcgacc ctatcgtgca gcaccggccc atgctggagg agaggatcgt
     2641 ggagctgttc agaaacttaa tgaagagtgc cttcgtggtg gagcggcagc cctgcatgcc
     2701 catgcacccg gaccggccct tagtcatcaa gactggtgtc cagtttacca cgaaagtcag
     2761 gttgctggtc aaatttcctg agttgaatta tcagcttaaa attaaagtgt gcattgataa
     2821 agactctggg gatgttgctg ccctcagagg gtctcggaaa tttaacattc tgggcacgaa
     2881 cacaaaagtg atgaacatgg aggagtctaa caacggcagc ctgtctgcag agttcaagca
     2941 cctgaccctt agggagcaga gatgtgggaa tggaggccgt gccaattgtg atgcctcctt
     3001 gatcgtgact gaggagctgc acctgatcac cttcgagact gaggtgtacc accaaggcct
     3061 caagattgac ctagagaccc actccttgcc agttgtggtg atctccaaca tctgtcagat
     3121 gccaaatgct tgggcatcaa tcctgtggta taacatgctg accaataacc ccaagaacgt
     3181 gaacttcttc actaagccgc caattggaac ctgggaccaa gtggccgagg tgctcagctg
     3241 gcagttctcg tccaccacca agcgggggct gagcatcgag cagctgacaa cgctggctga
     3301 gaagctccta gggcctggtg tgaactactc agggtgtcag atcacatggg ctaaattctg
     3361 caaagaaaac atggctggca agggcttctc cttctgggtc tggctagaca atatcatcga
     3421 ccttgtgaaa aagtatatct tggccctttg gaatgaaggg tacatcatgg gtttcatcag
     3481 caaggagcgg gagcgggcca tcctaagcac aaagcccccg ggcaccttcc tactgcagtt
     3541 cagcgagagc agcaaagaag gaggggtcac tttcacttgg gtggaaaagg acatcagtgg
     3601 caagacccag atccagtctg tagagccata caccaagcag cagctgaaca acatgtcatt
     3661 tgctgaaatc atcatgggct ataagatcat ggatgcgacc aacatcctgg tgtctccact
     3721 tgtctacctc taccccgaca ttcccaagga ggaggcattc ggaaagtact gtaggcccga
     3781 gagccaggag caccccgaag ccgacccagg tagtgctgcc ccgtacctga agaccaagtt
     3841 catctgtgtg acaccaacga cctgcagcaa taccattgac ctgccgatgt ccccccgcac
     3901 tttagattca ttgatgcagt ttggaaataa cggtgaaggt gctgagccct cagcaggagg
     3961 gcagtttgag tcgctcacgt ttgacatgga tctgacctcg gagtgtgcta cctcccccat
     4021 gtaagaattc agatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     4081 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     4141 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     4201 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     4261 gtagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     4321 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     4381 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     4441 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     4501 gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     4561 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     4621 aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4681 ggacggcgac gtaaacggcc acaagttcag cgtgaggggc gagggcgagg gcgatgccac
     4741 caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     4801 caccctcgtg accaccctga gccacggcgt gcagtgcttc gcccgctacc ccgaccacat
     4861 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     4921 cttcttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac
     4981 cctggtgaac cgcatcgagc tgaagggcgt cgacttcaag gaggacggca acatcctggg
     5041 gcacaagctg gagtacaact tcaacagcca caacatctat atcatggccg tcaagcagaa
     5101 gaacggcatc aaggtgaact tcaagatccg ccacaacgtg gaggacggca gcgtgcagct
     5161 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacag
     5221 ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat
     5281 ggtcctgctg gagttccgca ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     5341 gtaaaccggt gctcgaggta ccgatgaata gctaaggtag aggccgcagg taagtatcaa
     5401 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     5461 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     5521 aggtgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5581 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5641 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5701 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5761 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5821 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5881 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5941 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     6001 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     6061 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     6121 tggaattcga gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     6181 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     6241 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     6301 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcccagc ttttgttccc
     6361 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6421 attgttatcc gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     6481 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     6541 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     6601 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     6661 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     6721 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     6781 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     6841 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     6901 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     6961 tcggccggcc gccggcgtcg acgcgcgcgg ccgcggcgcg ccgttgctgg cgtttttcca
     7021 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7081 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7141 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7201 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7261 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7321 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7381 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7441 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7501 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7561 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7621 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7681 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7741 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7801 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7861 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7921 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7981 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8041 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8101 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8161 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8221 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8281 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8341 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8401 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8461 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8521 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8581 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8641 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8701 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8761 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     8821 gaggcccttt cgtctacgcg cgcgccggcg gcgcgccgtc gacggccggc cgcggccgc
//
