LOCUS       sequence-249375-        6299 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 6299)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6299)
  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-185    N-Tag GFP[2] L10 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6299
                     /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"
     CDS             1121..1444
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(1458..2032,2033..2035,2036..2044)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1625..1642)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1852..1871
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2045..2047,2048..2050,2051..2764)
                     /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(2090..2111)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2810..2942
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2951..3539
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3004..3024)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3422..3433)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3566..3583)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3572..3683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3779..3799)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3779..3797)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3818..3834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3818..3834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3831..3853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3936..3955)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4001..4056
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4055..4074)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4435..5023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4515..4534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5194..5985,5986..6054))
                     /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     5817..5836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6055..6159)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6227..6245
                     /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 caccatggaa aagagggacc acatggtgct gctcgagtac gtcacagcag ctggcatcac
     1081 cgatgccagc gggagctctg ccagcggtgg cagcacctcc ggagtgcagg tggaaaccat
     1141 ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg tgcactacac
     1201 cgggatgctt gaagatggaa agaaatttga ttcctcccgg gacagaaaca agccctttaa
     1261 gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg cccagatgag
     1321 tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg ccactgggca
     1381 cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc taaaactgga
     1441 ataagaattc agatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     1501 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     1561 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     1621 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     1681 gtagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     1741 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     1801 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     1861 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     1921 gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     1981 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     2041 aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2101 ggacggcgac gtaaacggcc acaagttcag cgtgaggggc gagggcgagg gcgatgccac
     2161 caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2221 caccctcgtg accaccctga gccacggcgt gcagtgcttc gcccgctacc ccgaccacat
     2281 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2341 cttcttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac
     2401 cctggtgaac cgcatcgagc tgaagggcgt cgacttcaag gaggacggca acatcctggg
     2461 gcacaagctg gagtacaact tcaacagcca caacatctat atcatggccg tcaagcagaa
     2521 gaacggcatc aaggtgaact tcaagatccg ccacaacgtg gaggacggca gcgtgcagct
     2581 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacag
     2641 ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat
     2701 ggtcctgctg gagttccgca ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2761 gtaaaccggt gctcgaggta ccgatgaata gctaaggtag aggccgcagg taagtatcaa
     2821 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     2881 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     2941 aggtgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3001 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3061 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3121 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3181 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3241 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3301 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3361 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3421 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3481 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3541 tggaattcga gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3601 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3661 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3721 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcccagc ttttgttccc
     3781 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3841 attgttatcc gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     3901 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     3961 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     4021 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     4081 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     4141 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     4201 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     4261 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     4321 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     4381 tcggccggcc gccggcgtcg acgcgcgcgg ccgcggcgcg ccgttgctgg cgtttttcca
     4441 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4501 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4561 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4621 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4681 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4741 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4801 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4861 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4921 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4981 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5041 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5101 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5161 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5221 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5281 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5341 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5401 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5461 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5521 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5581 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5641 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5701 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5761 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5821 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5881 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5941 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6001 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6061 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6121 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6181 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6241 gaggcccttt cgtctacgcg cgcgccggcg gcgcgccgtc gacggccggc cgcggccgc
//
