LOCUS       sequence-270703-        6185 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 6185)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6185)
  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-144    C-Tag GFP[2] L32 GCNZipper EBPF2
FEATURES             Location/Qualifiers
     source          1..6185
                     /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"
     CDS             1097..1153
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     misc_feature    join(1344..1918,1919..1921,1922..1930)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1511..1528)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1738..1757
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1931..1933,1934..1936,1937..2650)
                     /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(1976..1997)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2696..2828
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2837..3425
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2890..2910)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3308..3319)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3452..3469)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3458..3569
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3665..3685)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3665..3683)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3704..3720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3704..3720)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3717..3739)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3822..3841)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3887..3942
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3941..3960)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4321..4909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4401..4420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5080..5871,5872..5940))
                     /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     5703..5722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5941..6045)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6113..6131
                     /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 caccatgaac actgaagccg ccaggcgttc tcgtgcgaga aagttgcaaa gaatgaaaca
     1081 acttgaagac aaggttgaag aattgctttc gaaaaattat cacttggaaa atgaggttgc
     1141 cagattaaag aaattagttg gcgaacgcga cgtgggtggc ggagggagcg aaggcggcgg
     1201 atccggaggg cctggctctg gcggagaggg gagcgctgga ggcgggtcag ccggtggtgg
     1261 gagtgaaaag agggaccaca tggtgctgct cgagtacgtc acagcagctg gcatcaccga
     1321 tgccagctaa gaattcagat ccgcccctct ccctcccccc cccctaacgt tactggccga
     1381 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     1441 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     1501 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     1561 cctctggtag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     1621 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     1681 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     1741 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     1801 ggatctgatc tggggcctcg gtacacatgc tttacatgtg tttagtcgag gttaaaaaaa
     1861 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     1921 ggccacaacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1981 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg aggggcgagg gcgagggcga
     2041 tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2101 ctggcccacc ctcgtgacca ccctgagcca cggcgtgcag tgcttcgccc gctaccccga
     2161 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2221 caccatcttc ttcaaggacg acggcaccta caagacccgc gccgaggtga agttcgaggg
     2281 cgacaccctg gtgaaccgca tcgagctgaa gggcgtcgac ttcaaggagg acggcaacat
     2341 cctggggcac aagctggagt acaacttcaa cagccacaac atctatatca tggccgtcaa
     2401 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacgtggagg acggcagcgt
     2461 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2521 cgacagccac tacctgagca cccagtccgt gctgagcaaa gaccccaacg agaagcgcga
     2581 tcacatggtc ctgctggagt tccgcaccgc cgccgggatc actctcggca tggacgagct
     2641 gtacaagtaa accggtgctc gaggtaccga tgaatagcta aggtcgaggc cgcaggtaag
     2701 tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag
     2761 aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
     2821 ctccacaggt gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2881 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2941 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3001 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3061 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3121 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3181 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     3241 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3301 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3361 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3421 cccgcctgga attcgagctc ggtacgatca gcctcgactg tgccttctag ttgccagcca
     3481 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3541 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3601 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag cccagctttt
     3661 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     3721 tgtgaaattg ttatccgcta attcactcct caggtgcagg ctgcctatca gaaggtggtg
     3781 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
     3841 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     3901 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     3961 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     4021 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     4081 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     4141 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     4201 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     4261 tggagatcgg ccggccgccg gcgtcgacgc gcgcggccgc ggcgcgccgt tgctggcgtt
     4321 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4381 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4441 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4501 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4561 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4621 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4681 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4741 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4801 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4861 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4921 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4981 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5041 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5101 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5161 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5221 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5281 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5341 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5401 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5461 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5521 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5581 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5641 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5701 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5761 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5821 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5881 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5941 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6001 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6061 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6121 tatcacgagg ccctttcgtc tacgcgcgcg ccggcggcgc gccgtcgacg gccggccgcg
     6181 gccgc
//
