LOCUS       sequence-249270-        6638 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 6638)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6638)
  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-207    C-Tag miRFP670[1] L32 FRB EBPF2
FEATURES             Location/Qualifiers
     source          1..6638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     732..756
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          871..1003
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1018..1027
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1022..1031
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1033..1308
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRIS"
     misc_feature    join(1796..2370,2371..2373,2374..2382)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1963..1980)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2190..2209
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2383..2385,2386..2388,2389..3102)
                     /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(2428..2449)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3148..3280
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3289..3877
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3342..3362)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3760..3771)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3904..3921)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3910..4021
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4117..4137)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4117..4135)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4156..4172)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4156..4172)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4169..4191)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4274..4293)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4339..4394
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4393..4412)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4773..5361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4853..4872)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5532..6323,6324..6392))
                     /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     6155..6174
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6393..6497)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6565..6583
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctgcctcca ggcctcaata ttggccatta gccatattat tcattggtta tatagcataa
       61 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
      121 attggctcat gtccaatatg accgccatgt tggcattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt
      781 tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc
      841 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
      901 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
      961 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccaggcc
     1021 accatggtgg ccatcctctg gcatgagatg tggcatgaag gcctggaaga ggcatctcgt
     1081 ttgtactttg gggaaaggaa cgtgaaaggc atgtttgagg tgctggagcc cttgcatgct
     1141 atgatggaac ggggccccca gactctgaag gaaacatcct ttaatcaggc ctatggtcga
     1201 gatttaatgg aggcccaaga gtggtgcagg aagtacatga aatcagggaa tgtcaaggac
     1261 ctcacccaag cctgggacct ctattatcat gtgttccgac gaatctcaga cgtgggtggc
     1321 ggagggagcg aaggcggcgg atccggaggg cctggctctg gcggagaggg gagcgctgga
     1381 ggcgggtcag ccggtggtgg gagtgtggct gggcatgcta gcggttctcc tgcctttggc
     1441 accgctagtc actccaactg cgagcacgag gagatccatc ttgctgggag tatccagcct
     1501 cacggagccc ttctggtcgt tagcgaacac gatcaccgtg tgattcaggc aagcgcgaat
     1561 gcagccgagt tcctcaacct gggcagtgtg ctgggagttc ccttggccga aatcgacgga
     1621 gatctgctga tcaagattct gccccatctc gatccgacag ctgaaggcat gccagtagca
     1681 gttaggtgtc gcattgggaa tccgtccacc gagtattgcg gtctgatgca taggccacca
     1741 gaaggagggc tgattatcga gctggaaaga gcgggaccct aagaattcag atccgcccct
     1801 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     1861 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     1921 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     1981 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctggt agcttcttga agacaaacaa
     2041 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     2101 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     2161 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     2221 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat
     2281 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     2341 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag
     2401 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2461 aagttcagcg tgaggggcga gggcgagggc gatgccacca acggcaagct gaccctgaag
     2521 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgagc
     2581 cacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     2641 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcacc
     2701 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2761 aagggcgtcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc
     2821 aacagccaca acatctatat catggccgtc aagcagaaga acggcatcaa ggtgaacttc
     2881 aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2941 acccccatcg gcgacggccc cgtgctgctg cccgacagcc actacctgag cacccagtcc
     3001 gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttccgcacc
     3061 gccgccggga tcactctcgg catggacgag ctgtacaagt aaaccggtgc tcgaggtacc
     3121 gatgaatagc taaggtcgag gccgcaggta agtatcaagg ttacaagaca ggtttaagga
     3181 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     3241 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtcgacaa tcaacctctg
     3301 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3361 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3421 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3481 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3541 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3601 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3661 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     3721 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3781 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3841 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc tcggtacgat
     3901 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3961 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4021 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4081 gggaggattg ggaagacaat agcccagctt ttgttccctt tagtgagggt taattgcgcg
     4141 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc taattcactc
     4201 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     4261 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     4321 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     4381 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     4441 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     4501 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     4561 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     4621 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     4681 tactcccagt catagctgtc cctcttctct tatggagatc ggccggccgc cggcgtcgac
     4741 gcgcgcggcc gcggcgcgcc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4801 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4861 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4921 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4981 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5041 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5101 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5161 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5221 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5281 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5341 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5401 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5461 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5521 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5581 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5641 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5701 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5761 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5821 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5881 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5941 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6001 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6061 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6121 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6181 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6241 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6301 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6361 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6421 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6481 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6541 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctacgcgcg
     6601 cgccggcggc gcgccgtcga cggccggccg cggccgcg
//
