LOCUS       sequence-249281-        6641 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 6641)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6641)
  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-114    N-Tag miRFP670[1] L20 FKBP EBPF2
FEATURES             Location/Qualifiers
     source          1..6641
                     /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             1463..1786
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(1800..2374,2375..2377,2378..2386)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1967..1984)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2194..2213
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2387..2389,2390..2392,2393..3106)
                     /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(2432..2453)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3152..3284
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3293..3881
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3346..3366)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3764..3775)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3908..3925)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3914..4025
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4121..4141)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4121..4139)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4160..4176)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4160..4176)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4173..4195)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4278..4297)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4343..4398
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4397..4416)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4777..5365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4857..4876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5536..6327,6328..6396))
                     /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     6159..6178
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6397..6501)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6569..6587
                     /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 ccgggagtcc tggcggtgga tccggaggct ctgggagctc
     1441 tgccagcggt ggcagcacct ccggagtgca ggtggaaacc atctccccag gagacgggcg
     1501 caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg
     1561 aaagaaattt gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca
     1621 ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa
     1681 actgactata tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc
     1741 acatgccact ctcgtcttcg atgtggagct tctaaaactg gaataagaat tcagatccgc
     1801 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     1861 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     1921 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     1981 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggtagcttc ttgaagacaa
     2041 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     2101 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     2161 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     2221 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac
     2281 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     2341 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg
     2401 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2461 ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc accaacggca agctgaccct
     2521 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2581 gagccacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     2641 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2701 cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2761 gctgaagggc gtcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2821 cttcaacagc cacaacatct atatcatggc cgtcaagcag aagaacggca tcaaggtgaa
     2881 cttcaagatc cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc actaccagca
     2941 gaacaccccc atcggcgacg gccccgtgct gctgcccgac agccactacc tgagcaccca
     3001 gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttccg
     3061 caccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaaccg gtgctcgagg
     3121 taccgatgaa tagctaaggt agaggccgca ggtaagtatc aaggttacaa gacaggttta
     3181 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
     3241 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcg acaatcaacc
     3301 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3361 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3421 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3481 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3541 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3601 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3661 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     3721 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3781 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3841 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaattc gagctcggta
     3901 cgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3961 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     4021 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4081 aagggggagg attgggaaga caatagccca gcttttgttc cctttagtga gggttaattg
     4141 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctaattc
     4201 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct
     4261 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc
     4321 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
     4381 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag
     4441 aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa
     4501 ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc
     4561 catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt
     4621 tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc
     4681 tgactactcc cagtcatagc tgtccctctt ctcttatgga gatcggccgg ccgccggcgt
     4741 cgacgcgcgc ggccgcggcg cgccgttgct ggcgtttttc cataggctcc gcccccctga
     4801 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4861 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4921 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4981 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5041 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5101 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5161 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5221 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5281 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5341 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5401 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5461 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5521 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5581 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5641 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5701 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5761 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5821 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5881 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5941 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6001 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6061 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6121 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6181 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6241 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6301 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6361 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6421 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6481 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6541 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctacg
     6601 cgcgcgccgg cggcgcgccg tcgacggccg gccgcggccg c
//
