LOCUS       sequence-264960-        6216 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 6216)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6216)
  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-339    C-Tag UnaG[1] L20 {Esp3I(2x)} EBPF2
FEATURES             Location/Qualifiers
     source          1..6216
                     /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"
     misc_feature    join(1374..1948,1949..1951,1952..1960)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1541..1558)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1768..1787
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1961..1963,1964..1966,1967..2680)
                     /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(2006..2027)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2726..2858
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2867..3455
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2920..2940)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3338..3349)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3482..3499)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3488..3599
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3695..3715)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3695..3713)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3734..3750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3734..3750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3747..3769)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3852..3871)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3917..3972
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3971..3990)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4351..4939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4431..4450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5110..5901,5902..5970))
                     /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     5733..5752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5971..6075)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6143..6161
                     /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 acccgagacg ccgaggcacg ccgtctccgg gagtcctggc ggtggatccg gaggctctgg
     1081 gagctctgcc agcggtggca gcaccagtgt cgaaaagttc gtgggcacat ggaagattgc
     1141 cgatagccac aacttcggcg aatacctcaa agctataggg gcaccaaaag agcttagcga
     1201 tggtggagat gcgacaactc ccacactgta cattagccag aaagacggag acaagatgac
     1261 tgtcaagatc gagaatgggc ctcccacctt tctggacaca caggttaagt tcaaactggg
     1321 tgaggagttt gacgagtttc ccagtgatcg caggaaataa gaattcagat ccgcccctct
     1381 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     1441 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     1501 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     1561 ggtctgttga atgtcgtgaa ggaagcagtt cctctggtag cttcttgaag acaaacaacg
     1621 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     1681 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     1741 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     1801 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtacacatgc
     1861 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     1921 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga
     1981 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2041 gttcagcgtg aggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     2101 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca
     2161 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2221 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaccta
     2281 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2341 gggcgtcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
     2401 cagccacaac atctatatca tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa
     2461 gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2521 ccccatcggc gacggccccg tgctgctgcc cgacagccac tacctgagca cccagtccgt
     2581 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tccgcaccgc
     2641 cgccgggatc actctcggca tggacgagct gtacaagtaa accggtgctc gaggtaccga
     2701 tgaatagcta aggtcgaggc cgcaggtaag tatcaaggtt acaagacagg tttaaggaga
     2761 ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat
     2821 tggtcttact gacatccact ttgcctttct ctccacaggt gtcgacaatc aacctctgga
     2881 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2941 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3001 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3061 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3121 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3181 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3241 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3301 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3361 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3421 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc ggtacgatca
     3481 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3541 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3601 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3661 gaggattggg aagacaatag cccagctttt gttcccttta gtgagggtta attgcgcgct
     3721 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgcta attcactcct
     3781 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     3841 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     3901 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     3961 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     4021 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     4081 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     4141 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     4201 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     4261 ctcccagtca tagctgtccc tcttctctta tggagatcgg ccggccgccg gcgtcgacgc
     4321 gcgcggccgc ggcgcgccgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4381 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4441 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4501 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4561 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4621 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4681 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4741 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4801 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4861 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4921 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4981 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5041 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5101 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5161 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5221 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5281 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5341 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5401 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5461 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5521 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5581 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5641 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5701 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5761 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5821 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5881 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5941 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6001 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6061 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6121 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tacgcgcgcg
     6181 ccggcggcgc gccgtcgacg gccggccgcg gccgcg
//
