LOCUS       sequence-249283-        6677 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 6677)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6677)
  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-116    N-Tag miRFP670[1] L32 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6677
                     /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"
     regulatory      1023..1032
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1499..1822
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(1836..2410,2411..2413,2414..2422)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2003..2020)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2230..2249
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2423..2425,2426..2428,2429..3142)
                     /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(2468..2489)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3188..3320
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3329..3917
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3382..3402)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3800..3811)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3944..3961)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3950..4061
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4157..4177)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4157..4175)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4196..4212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4196..4212)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4209..4231)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4314..4333)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4379..4434
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4433..4452)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4813..5401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4893..4912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5572..6363,6364..6432))
                     /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     6195..6214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6433..6537)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6605..6623
                     /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 gctgggcatg ctagcggttc tcctgccttt ggcaccgcta gtcactccaa
     1081 ctgcgagcac gaggagatcc atcttgctgg gagtatccag cctcacggag cccttctggt
     1141 cgttagcgaa cacgatcacc gtgtgattca ggcaagcgcg aatgcagccg agttcctcaa
     1201 cctgggcagt gtgctgggag ttcccttggc cgaaatcgac ggagatctgc tgatcaagat
     1261 tctgccccat ctcgatccga cagctgaagg catgccagta gcagttaggt gtcgcattgg
     1321 gaatccgtcc accgagtatt gcggtctgat gcataggcca ccagaaggag ggctgattat
     1381 cgagctggaa agagcgggac ccgacgtggg tggcggaggg agcgaaggcg gcggatccgg
     1441 agggcctggc tctggcggag aggggagcgc tggaggcggg tcagccggtg gtgggagtgg
     1501 agtgcaggtg gaaaccatct ccccaggaga cgggcgcacc ttccccaagc gcggccagac
     1561 ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaatttgatt cctcccggga
     1621 cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag gctgggaaga
     1681 aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc cagattatgc
     1741 ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg tcttcgatgt
     1801 ggagcttcta aaactggaat aagaattcag atccgcccct ctccctcccc cccccctaac
     1861 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     1921 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     1981 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     2041 aaggaagcag ttcctctggt agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     2101 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     2161 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     2221 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     2281 ccccattgta tgggatctga tctggggcct cggtacacat gctttacatg tgtttagtcg
     2341 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     2401 cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt
     2461 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgaggggcga
     2521 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2581 gctgcccgtg ccctggccca ccctcgtgac caccctgagc cacggcgtgc agtgcttcgc
     2641 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2701 cgtccaggag cgcaccatct tcttcaagga cgacggcacc tacaagaccc gcgccgaggt
     2761 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcgtcg acttcaagga
     2821 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acatctatat
     2881 catggccgtc aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacgtgga
     2941 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3001 cgtgctgctg cccgacagcc actacctgag cacccagtcc gtgctgagca aagaccccaa
     3061 cgagaagcgc gatcacatgg tcctgctgga gttccgcacc gccgccggga tcactctcgg
     3121 catggacgag ctgtacaagt aaaccggtgc tcgaggtacc gatgaatagc taaggtcgag
     3181 gccgcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
     3241 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
     3301 ctttgccttt ctctccacag gtgtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     3361 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3421 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3481 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3541 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3601 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3661 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3721 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3781 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3841 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3901 ttgggccgcc tccccgcctg gaattcgagc tcggtacgat cagcctcgac tgtgccttct
     3961 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     4021 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     4081 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     4141 agcccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     4201 agctgtttcc tgtgtgaaat tgttatccgc taattcactc ctcaggtgca ggctgcctat
     4261 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     4321 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     4381 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     4441 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     4501 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     4561 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     4621 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     4681 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     4741 cctcttctct tatggagatc ggccggccgc cggcgtcgac gcgcgcggcc gcggcgcgcc
     4801 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4861 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4921 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4981 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5041 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5101 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5161 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5221 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5281 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5341 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5401 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5461 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5521 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5641 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5701 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5761 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5821 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5881 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5941 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6001 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6061 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6121 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6181 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6241 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6301 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6361 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6421 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6481 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6541 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6601 taaaaatagg cgtatcacga ggccctttcg tctacgcgcg cgccggcggc gcgccgtcga
     6661 cggccggccg cggccgc
//
