LOCUS       sequence-249214-        6549 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 6549)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6549)
  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-333    C-Tag miRFP670[2] L20 {Esp3I(2x)}...
FEATURES             Location/Qualifiers
     source          1..6549
                     /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"
     misc_feature    join(1708..2282,2283..2285,2286..2294)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1875..1892)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2102..2121
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2295..2297,2298..2300,2301..3014)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2295..2297,2298..2300,2301..3011)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2340..2361)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2601..2620)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2948..2969
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3060..3192
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3201..3789
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3254..3274)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3672..3683)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3816..3833)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3822..3933
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4029..4049)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4029..4047)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4068..4084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4068..4084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4081..4103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4186..4205)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4251..4306
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4305..4324)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4685..5273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4765..4784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5444..6235,6236..6304))
                     /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     6067..6086
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6305..6409)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6477..6495
                     /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 cacccgagac gccgaggcac gccgtctccg ggagtcctgg cggtggatcc ggaggctctg
     1081 ggagctctgc cagcggtggc agcaccagtg gcagagcggg acccagtatc gacctgtctg
     1141 ggacactcgc accagcactg gagcgcataa gaactgctgg atcactgagg gccctctgtg
     1201 acgacacagt cttgctcttt cagcaatgta cgggctatga tcgagtgatg gtgtaccgat
     1261 tcgacgaaca gggtcatggt ctggtctttt ccgagtgtca cgtgcctggc ctggagagct
     1321 actttggcaa ccgctatccc tcttctaccg taccccagat ggccaggcaa ctttacgtcc
     1381 ggcaacgggt cagagtgctg gtggatgtga cctaccagcc tgtgccactc gaacctcggc
     1441 tgagcccact cactggacgg gatctggaca tgagcggctg cttcctgagg tccatgtcac
     1501 cttgccatct gcagttcttg aaggacatgg gcgttcgagc cacactcgca gtctcactgg
     1561 ttgtaggcgg gaaactgtgg ggtcttgtgg tgtgtcacca ctaccttccc aggttcatcc
     1621 ggttcgagtt gagagccatc tgcaaacgtt tggccgagag aatagccacc cgcattactg
     1681 ctctggaatc ctaagaattc agatccgccc ctctccctcc ccccccccta acgttactgg
     1741 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     1801 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     1861 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     1921 agttcctctg gtagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     1981 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     2041 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     2101 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     2161 tatgggatct gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa
     2221 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     2281 atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2341 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2401 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2461 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2521 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2581 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2641 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2701 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2761 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2821 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2881 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2941 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3001 agctgtacaa gtaaaccggt gctcgaggta ccgatgaata gctaaggtcg aggccgcagg
     3061 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     3121 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     3181 ttctctccac aggtgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     3241 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3301 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3361 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3421 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3481 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3541 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     3601 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3661 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3721 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3781 cctccccgcc tggaattcga gctcggtacg atcagcctcg actgtgcctt ctagttgcca
     3841 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3901 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3961 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcccagc
     4021 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     4081 cctgtgtgaa attgttatcc gctaattcac tcctcaggtg caggctgcct atcagaaggt
     4141 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     4201 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     4261 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     4321 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     4381 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     4441 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     4501 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     4561 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     4621 cttatggaga tcggccggcc gccggcgtcg acgcgcgcgg ccgcggcgcg ccgttgctgg
     4681 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4741 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4801 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4861 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4921 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4981 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5041 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5101 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5161 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5221 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5281 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5341 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5401 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5461 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5521 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5581 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5641 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5701 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5761 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5821 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5881 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5941 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6001 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6061 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6121 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6181 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6241 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6301 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6361 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6421 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     6481 ggcgtatcac gaggcccttt cgtctacgcg cgcgccggcg gcgcgccgtc gacggccggc
     6541 cgcggccgc
//
