LOCUS       sequence-249246-        6630 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 6630)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6630)
  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-365    C-Tag Venus210[1] L32 {Esp3I(2x)}...
FEATURES             Location/Qualifiers
     source          1..6630
                     /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(1788..2362,2363..2365,2366..2374)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1955..1972)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2182..2201
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2375..2377,2378..2380,2381..3094)
                     /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(2420..2441)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3140..3272
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3281..3869
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3334..3354)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3752..3763)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3896..3913)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3902..4013
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4109..4129)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4109..4127)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4148..4164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4148..4164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4161..4183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4266..4285)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4331..4386
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4385..4404)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4765..5353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4845..4864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5524..6315,6316..6384))
                     /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     6147..6166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6385..6489)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6557..6575
                     /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 ccgtctccga cgtgggtggc ggagggagcg aaggcggcgg
     1081 atccggaggg cctggctctg gcggagaggg gagcgctgga ggcgggtcag ccggtggtgg
     1141 cagtgtgtca aagggagaag agttgtttac cggtgttgtt cccatactgg tggaactgga
     1201 tggtgacgtc aatggacaca agtttagcgt ttctggcgaa ggagaaggcg atgctaccta
     1261 tggcaaactg accctcaaac tgatctgtac gactggaaag ttgcctgtac cttggccaac
     1321 actggtcaca accttgggct atgggctgca atgctttgcc cgttatcctg atcacatgaa
     1381 gcaacatgac ttctttaagt ctgctatgcc agagggatat gtgcaggaac ggactatctt
     1441 cttcaaggac gacggcaact acaagacgag agctgaggtg aaatttgagg gtgacacact
     1501 ggtgaatcgc atagagctga aaggcatcga cttcaaagaa gatgggaata ttctcgggca
     1561 taaactcgag tacaactaca attcccacaa cgtgtacatc actgccgata agcagaagaa
     1621 tgggattaag gcgaacttca agatccgaca caacatcgag gatggtgggg tccagcttgc
     1681 cgaccattat cagcagaaca cacccattgg ggatggacca gtcctgttgc cggataacca
     1741 ctacctcagt taccagagca aactgtccaa ataagaattc agatccgccc ctctccctcc
     1801 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     1861 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     1921 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     1981 ttgaatgtcg tgaaggaagc agttcctctg gtagcttctt gaagacaaac aacgtctgta
     2041 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     2101 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     2161 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     2221 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     2281 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     2341 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt
     2401 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2461 cgtgaggggc gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg
     2521 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga gccacggcgt
     2581 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2641 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca cctacaagac
     2701 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcgt
     2761 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca
     2821 caacatctat atcatggccg tcaagcagaa gaacggcatc aaggtgaact tcaagatccg
     2881 ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2941 cggcgacggc cccgtgctgc tgcccgacag ccactacctg agcacccagt ccgtgctgag
     3001 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttccgca ccgccgccgg
     3061 gatcactctc ggcatggacg agctgtacaa gtaaaccggt gctcgaggta ccgatgaata
     3121 gctaaggtcg aggccgcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
     3181 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
     3241 tactgacatc cactttgcct ttctctccac aggtgtcgac aatcaacctc tggattacaa
     3301 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3361 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3421 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3481 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3541 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3601 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3661 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3721 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3781 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3841 tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacg atcagcctcg
     3901 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3961 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4021 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4081 tgggaagaca atagcccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     4141 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctaattcac tcctcaggtg
     4201 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     4261 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     4321 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     4381 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     4441 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     4501 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     4561 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     4621 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     4681 gtcatagctg tccctcttct cttatggaga tcggccggcc gccggcgtcg acgcgcgcgg
     4741 ccgcggcgcg ccgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5341 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5461 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5521 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5581 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5641 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5701 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5761 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5821 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5881 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5941 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6001 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6061 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6121 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6181 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6241 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6301 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6361 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6421 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6481 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     6541 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctacgcg cgcgccggcg
     6601 gcgcgccgtc gacggccggc cgcggccgcg
//
