LOCUS       sequence-249231-        6456 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 6456)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6456)
  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-350 N-Tag Venus173/155[1] L10 {Esp3I(2x)...
FEATURES             Location/Qualifiers
     source          1..6456
                     /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(1615..2189,2190..2192,2193..2201)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1782..1799)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2009..2028
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2202..2204,2205..2207,2208..2921)
                     /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(2247..2268)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2967..3099
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3108..3696
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3161..3181)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3579..3590)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3723..3740)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3729..3840
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3936..3956)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3936..3954)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3975..3991)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3975..3991)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3988..4010)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4093..4112)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4158..4213
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4212..4231)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4592..5180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4672..4691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5351..6142,6143..6211))
                     /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     5974..5993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6212..6316)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6384..6402
                     /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 gaggggagct ctgccagcgg
     1561 tggcagcacc tcccgagacg ccgaggcacg ccgtctccta agaattcaga tccgcccctc
     1621 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     1681 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     1741 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     1801 aggtctgttg aatgtcgtga aggaagcagt tcctctggta gcttcttgaa gacaaacaac
     1861 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     1921 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     1981 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     2041 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtacacatg
     2101 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     2161 ggttttcctt tgaaaaacac gatgataata tggccacaac catggtgagc aagggcgagg
     2221 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2281 agttcagcgt gaggggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagt
     2341 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgagcc
     2401 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2461 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcacct
     2521 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2581 agggcgtcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacttca
     2641 acagccacaa catctatatc atggccgtca agcagaagaa cggcatcaag gtgaacttca
     2701 agatccgcca caacgtggag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2761 cccccatcgg cgacggcccc gtgctgctgc ccgacagcca ctacctgagc acccagtccg
     2821 tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttccgcaccg
     2881 ccgccgggat cactctcggc atggacgagc tgtacaagta aaccggtgct cgaggtaccg
     2941 atgaatagct aaggtagagg ccgcaggtaa gtatcaaggt tacaagacag gtttaaggag
     3001 accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta
     3061 ttggtcttac tgacatccac tttgcctttc tctccacagg tgtcgacaat caacctctgg
     3121 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3181 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3241 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3301 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3361 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3421 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3481 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     3541 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3601 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3661 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattcgagct cggtacgatc
     3721 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3781 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3841 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3901 ggaggattgg gaagacaata gcccagcttt tgttcccttt agtgagggtt aattgcgcgc
     3961 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct aattcactcc
     4021 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     4081 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     4141 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     4201 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     4261 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     4321 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     4381 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     4441 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     4501 actcccagtc atagctgtcc ctcttctctt atggagatcg gccggccgcc ggcgtcgacg
     4561 cgcgcggccg cggcgcgccg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4621 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4681 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4741 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4801 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4861 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4921 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4981 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5041 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5101 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5161 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5221 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5281 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5341 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5401 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5461 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5521 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5581 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5641 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5701 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5761 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5821 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5881 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5941 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6001 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6061 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6121 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6181 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6241 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6301 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     6361 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctacgcgcgc
     6421 gccggcggcg cgccgtcgac ggccggccgc ggccgc
//
