LOCUS       sequence-249221-        6219 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 6219)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6219)
  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-340    N-Tag UnaG[1] L20 {Esp3I(2x)} EBPF2
FEATURES             Location/Qualifiers
     source          1..6219
                     /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(1378..1952,1953..1955,1956..1964)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1545..1562)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1772..1791
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1965..1967,1968..1970,1971..2684)
                     /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(2010..2031)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2730..2862
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2871..3459
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2924..2944)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3342..3353)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3486..3503)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3492..3603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3699..3719)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3699..3717)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3738..3754)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3738..3754)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3751..3773)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3856..3875)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3921..3976
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3975..3994)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4355..4943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4435..4454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5114..5905,5906..5974))
                     /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     5737..5756
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5975..6079)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6147..6165
                     /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 caccatggtc gaaaagttcg tgggcacatg gaagattgcc gatagccaca acttcggcga
     1081 atacctcaaa gctatagggg caccaaaaga gcttagcgat ggtggagatg cgacaactcc
     1141 cacactgtac attagccaga aagacggaga caagatgact gtcaagatcg agaatgggcc
     1201 tcccaccttt ctggacacac aggttaagtt caaactgggt gaggagtttg acgagtttcc
     1261 cagtgatcgc aggaaaggga gtcctggcgg tggatccgga ggctctggga gctctgccag
     1321 cggtggcagc acctcccgag acgccgaggc acgccgtctc ctaagaattc agatccgccc
     1381 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     1441 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     1501 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     1561 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gtagcttctt gaagacaaac
     1621 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     1681 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     1741 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     1801 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac
     1861 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     1921 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg
     1981 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2041 acaagttcag cgtgaggggc gagggcgagg gcgatgccac caacggcaag ctgaccctga
     2101 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2161 gccacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     2221 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2281 cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2341 tgaagggcgt cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2401 tcaacagcca caacatctat atcatggccg tcaagcagaa gaacggcatc aaggtgaact
     2461 tcaagatccg ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2521 acacccccat cggcgacggc cccgtgctgc tgcccgacag ccactacctg agcacccagt
     2581 ccgtgctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttccgca
     2641 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaaccggt gctcgaggta
     2701 ccgatgaata gctaaggtag aggccgcagg taagtatcaa ggttacaaga caggtttaag
     2761 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     2821 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtcgac aatcaacctc
     2881 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2941 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3001 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3061 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3121 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3181 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3241 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     3301 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3361 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     3421 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacg
     3481 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3541 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3601 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3661 gggggaggat tgggaagaca atagcccagc ttttgttccc tttagtgagg gttaattgcg
     3721 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctaattcac
     3781 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     3841 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     3901 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3961 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     4021 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     4081 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     4141 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     4201 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     4261 actactccca gtcatagctg tccctcttct cttatggaga tcggccggcc gccggcgtcg
     4321 acgcgcgcgg ccgcggcgcg ccgttgctgg cgtttttcca taggctccgc ccccctgacg
     4381 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4441 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4501 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4561 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4621 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4681 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4741 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4801 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4861 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4921 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4981 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5041 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5101 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5161 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5221 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5281 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5341 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5401 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5461 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5521 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5581 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5641 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5701 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5761 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5821 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5881 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5941 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6001 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6061 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6121 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctacgcg
     6181 cgcgccggcg gcgcgccgtc gacggccggc cgcggccgc
//
