LOCUS       sequence-249258-        6066 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 6066)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6066)
  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-128    C-Tag GFP[2] L32 {Esp3I(2x)} EBPF2
FEATURES             Location/Qualifiers
     source          1..6066
                     /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"
     misc_feature    join(1225..1799,1800..1802,1803..1811)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1392..1409)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1619..1638
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1812..1814,1815..1817,1818..2531)
                     /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(1857..1878)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2577..2709
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2718..3306
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2771..2791)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3189..3200)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3333..3350)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3339..3450
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3546..3566)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3546..3564)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3585..3601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3585..3601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3598..3620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3703..3722)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3768..3823
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3822..3841)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4202..4790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4282..4301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4961..5752,5753..5821))
                     /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     5584..5603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5822..5926)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5994..6012
                     /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 cacccgagac gccgaggcac gccgtctccg acgtgggtgg cggagggagc gaaggcggcg
     1081 gatccggagg gcctggctct ggcggagagg ggagcgctgg aggcgggtca gccggtggtg
     1141 ggagtgaaaa gagggaccac atggtgctgc tcgagtacgt cacagcagct ggcatcaccg
     1201 atgccagcta agaattcaga tccgcccctc tccctccccc ccccctaacg ttactggccg
     1261 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     1321 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     1381 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     1441 tcctctggta gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     1501 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     1561 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     1621 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     1681 gggatctgat ctggggcctc ggtacacatg ctttacatgt gtttagtcga ggttaaaaaa
     1741 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     1801 tggccacaac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1861 tcgagctgga cggcgacgta aacggccaca agttcagcgt gaggggcgag ggcgagggcg
     1921 atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1981 cctggcccac cctcgtgacc accctgagcc acggcgtgca gtgcttcgcc cgctaccccg
     2041 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2101 gcaccatctt cttcaaggac gacggcacct acaagacccg cgccgaggtg aagttcgagg
     2161 gcgacaccct ggtgaaccgc atcgagctga agggcgtcga cttcaaggag gacggcaaca
     2221 tcctggggca caagctggag tacaacttca acagccacaa catctatatc atggccgtca
     2281 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacgtggag gacggcagcg
     2341 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2401 ccgacagcca ctacctgagc acccagtccg tgctgagcaa agaccccaac gagaagcgcg
     2461 atcacatggt cctgctggag ttccgcaccg ccgccgggat cactctcggc atggacgagc
     2521 tgtacaagta aaccggtgct cgaggtaccg atgaatagct aaggtcgagg ccgcaggtaa
     2581 gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga
     2641 gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc
     2701 tctccacagg tgtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2761 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2821 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2881 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2941 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3001 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3061 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     3121 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     3181 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3241 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3301 ccccgcctgg aattcgagct cggtacgatc agcctcgact gtgccttcta gttgccagcc
     3361 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3421 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3481 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcccagcttt
     3541 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     3601 gtgtgaaatt gttatccgct aattcactcc tcaggtgcag gctgcctatc agaaggtggt
     3661 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     3721 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     3781 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     3841 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     3901 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     3961 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     4021 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     4081 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     4141 atggagatcg gccggccgcc ggcgtcgacg cgcgcggccg cggcgcgccg ttgctggcgt
     4201 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4261 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4321 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4381 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4441 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4501 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4561 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4621 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4681 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5941 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     6001 gtatcacgag gccctttcgt ctacgcgcgc gccggcggcg cgccgtcgac ggccggccgc
     6061 ggccgc
//
