LOCUS       sequence-249379-        6365 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 6365)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6365)
  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-252    N-Tag GFP[2] L32 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6365
                     /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             1187..1510
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(1524..2098,2099..2101,2102..2110)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1691..1708)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1918..1937
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2111..2113,2114..2116,2117..2830)
                     /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(2156..2177)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2876..3008
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3017..3605
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3070..3090)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3488..3499)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3632..3649)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3638..3749
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3845..3865)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3845..3863)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3884..3900)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3884..3900)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3897..3919)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4002..4021)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4067..4122
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4121..4140)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4501..5089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4581..4600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5260..6051,6052..6120))
                     /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     5883..5902
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6121..6225)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6293..6311
                     /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 gacgtgggtg gcggagggag cgaaggcggc ggatccggag ggcctggctc
     1141 tggcggagag gggagcgctg gaggcgggtc agccggtggt gggagtggag tgcaggtgga
     1201 aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca
     1261 ctacaccggg atgcttgaag atggaaagaa atttgattcc tcccgggaca gaaacaagcc
     1321 ctttaagttt atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca
     1381 gatgagtgtg ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac
     1441 tgggcaccca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa
     1501 actggaataa gaattcagat ccgcccctct ccctcccccc cccctaacgt tactggccga
     1561 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     1621 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     1681 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     1741 cctctggtag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     1801 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     1861 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     1921 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     1981 ggatctgatc tggggcctcg gtacacatgc tttacatgtg tttagtcgag gttaaaaaaa
     2041 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     2101 ggccacaacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2161 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg aggggcgagg gcgagggcga
     2221 tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2281 ctggcccacc ctcgtgacca ccctgagcca cggcgtgcag tgcttcgccc gctaccccga
     2341 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2401 caccatcttc ttcaaggacg acggcaccta caagacccgc gccgaggtga agttcgaggg
     2461 cgacaccctg gtgaaccgca tcgagctgaa gggcgtcgac ttcaaggagg acggcaacat
     2521 cctggggcac aagctggagt acaacttcaa cagccacaac atctatatca tggccgtcaa
     2581 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacgtggagg acggcagcgt
     2641 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2701 cgacagccac tacctgagca cccagtccgt gctgagcaaa gaccccaacg agaagcgcga
     2761 tcacatggtc ctgctggagt tccgcaccgc cgccgggatc actctcggca tggacgagct
     2821 gtacaagtaa accggtgctc gaggtaccga tgaatagcta aggtcgaggc cgcaggtaag
     2881 tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag
     2941 aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
     3001 ctccacaggt gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3061 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3121 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3181 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3241 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3301 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3361 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     3421 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3481 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3541 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3601 cccgcctgga attcgagctc ggtacgatca gcctcgactg tgccttctag ttgccagcca
     3661 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3721 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3781 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag cccagctttt
     3841 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     3901 tgtgaaattg ttatccgcta attcactcct caggtgcagg ctgcctatca gaaggtggtg
     3961 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
     4021 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     4081 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     4141 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     4201 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     4261 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     4321 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     4381 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     4441 tggagatcgg ccggccgccg gcgtcgacgc gcgcggccgc ggcgcgccgt tgctggcgtt
     4501 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4561 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4621 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4681 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4741 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4801 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4861 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4921 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4981 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5041 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5101 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5161 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5221 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5281 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5341 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5401 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5461 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5521 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5581 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5641 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5701 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5761 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5821 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5881 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5941 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6001 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6061 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6121 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6181 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6241 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6301 tatcacgagg ccctttcgtc tacgcgcgcg ccggcggcgc gccgtcgacg gccggccgcg
     6361 gccgc
//
