LOCUS       sequence-249402-        7691 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 7691)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7691)
  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-545    C-Tag Venus210[1] L32 Tln1(rod) E...
FEATURES             Location/Qualifiers
     source          1..7691
                     /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"
     misc_feature    join(2849..3423,3424..3426,3427..3435)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3016..3033)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3243..3262
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3436..3438,3439..3441,3442..4155)
                     /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(3481..3502)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          4201..4333
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    4342..4930
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4395..4415)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4813..4824)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4957..4974)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4963..5074
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5170..5190)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5170..5188)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5209..5225)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5209..5225)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5222..5244)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(5327..5346)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5392..5447
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5446..5465)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5826..6414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5906..5925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6585..7376,7377..7445))
                     /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     7208..7227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7446..7550)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7618..7636
                     /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 accatggctg tgtctggtat cattgctgac ctcgacacca ccatcatgtt cgctactgct
     1081 ggcacactta accgtgaggg tgctgaaact tttgctgacc accgggaggg tatcttaaag
     1141 acagcaaagg ttcttgtgga ggacaccaag gtcctagtgc agaatgcagc tgggagccag
     1201 gagaagttgg cacaagccgc ccagtcctcc gtggccacta ttacccgcct cgctgatgtg
     1261 gtcaagctcg gtgcagccag cctaggagcc gaagaccctg aaactcaggt ggtgctgatc
     1321 aatgcagtaa aggacgtagc caaggccctg ggtgacctca tcagcgctac gaaggctgca
     1381 gcgggcaaag ttggggatga ccctgcagtg tggcagctca agaactctgc caaggtgatg
     1441 gtgaccaatg tgacatcatt gctcaagaca gtgaaggctg tggaagatga ggccaccaaa
     1501 ggcacacggg ccctagaggc aaccacagag cacatacgtc aggaactggc ggtcttctgt
     1561 tccccagagc cacctgccaa gacctctacc cctgaagact tcatccgaat gaccaagggt
     1621 attactatgg caacagccaa agccgttgct gctggcaatt cctgtcgaca ggaagatgtc
     1681 attgccacag ccaatctgag ccgacgggct attgcggaca tgcttcgggc ttgcaaggaa
     1741 gcagctttcc acccagaagt ggcgcctgat gtacggctcc gagccctgca ctatggccgg
     1801 gagtgtgcca atggttacct ggaactgctg gaccacgtgc tgctgaccct tcagaagcca
     1861 aacccagacc tgaagcagca gctaactgga cactcaaagc gggttgctgg ctctgtgact
     1921 gagctcatcc aggctgctga agccatgaaa ggaacagagt gggtggaccc agaggaccct
     1981 actgtcattg ctgagaatga gctcttggga gccgcagccg ccatcgaggc tgcagccaag
     2041 aagctggagc agctgaagcc cagggccaaa cccaaggagg cggatgagtc cttgaacttt
     2101 gaggaacaag acgtgggtgg cggagggagc gaaggcggcg gatccggagg gcctggctct
     2161 ggcggagagg ggagcgctgg aggcgggtca gccggtggtg gcagtgtgtc aaagggagaa
     2221 gagttgttta ccggtgttgt tcccatactg gtggaactgg atggtgacgt caatggacac
     2281 aagtttagcg tttctggcga aggagaaggc gatgctacct atggcaaact gaccctcaaa
     2341 ctgatctgta cgactggaaa gttgcctgta ccttggccaa cactggtcac aaccttgggc
     2401 tatgggctgc aatgctttgc ccgttatcct gatcacatga agcaacatga cttctttaag
     2461 tctgctatgc cagagggata tgtgcaggaa cggactatct tcttcaagga cgacggcaac
     2521 tacaagacga gagctgaggt gaaatttgag ggtgacacac tggtgaatcg catagagctg
     2581 aaaggcatcg acttcaaaga agatgggaat attctcgggc ataaactcga gtacaactac
     2641 aattcccaca acgtgtacat cactgccgat aagcagaaga atgggattaa ggcgaacttc
     2701 aagatccgac acaacatcga ggatggtggg gtccagcttg ccgaccatta tcagcagaac
     2761 acacccattg gggatggacc agtcctgttg ccggataacc actacctcag ttaccagagc
     2821 aaactgtcca aataagaatt cagatccgcc cctctccctc ccccccccct aacgttactg
     2881 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     2941 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     3001 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     3061 cagttcctct ggtagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     3121 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     3181 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     3241 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     3301 gtatgggatc tgatctgggg cctcggtaca catgctttac atgtgtttag tcgaggttaa
     3361 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     3421 aatatggcca caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3481 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgagggg cgagggcgag
     3541 ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3601 gtgccctggc ccaccctcgt gaccaccctg agccacggcg tgcagtgctt cgcccgctac
     3661 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3721 gagcgcacca tcttcttcaa ggacgacggc acctacaaga cccgcgccga ggtgaagttc
     3781 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggcg tcgacttcaa ggaggacggc
     3841 aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacatcta tatcatggcc
     3901 gtcaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacgt ggaggacggc
     3961 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4021 ctgcccgaca gccactacct gagcacccag tccgtgctga gcaaagaccc caacgagaag
     4081 cgcgatcaca tggtcctgct ggagttccgc accgccgccg ggatcactct cggcatggac
     4141 gagctgtaca agtaaaccgg tgctcgaggt accgatgaat agctaaggtc gaggccgcag
     4201 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
     4261 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
     4321 tttctctcca caggtgtcga caatcaacct ctggattaca aaatttgtga aagattgact
     4381 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4441 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4501 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4561 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4621 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4681 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     4741 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     4801 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4861 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4921 gcctccccgc ctggaattcg agctcggtac gatcagcctc gactgtgcct tctagttgcc
     4981 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5041 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5101 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcccag
     5161 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     5221 tcctgtgtga aattgttatc cgctaattca ctcctcaggt gcaggctgcc tatcagaagg
     5281 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     5341 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     5401 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     5461 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     5521 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     5581 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     5641 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     5701 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     5761 tcttatggag atcggccggc cgccggcgtc gacgcgcgcg gccgcggcgc gccgttgctg
     5821 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5881 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5941 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6001 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6061 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6121 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6181 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6241 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6301 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6361 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6421 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6481 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6541 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6601 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6661 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6721 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6781 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6841 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6901 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6961 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7021 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7081 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7141 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7201 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7261 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7321 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7381 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7441 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7501 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7561 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     7621 aggcgtatca cgaggccctt tcgtctacgc gcgcgccggc ggcgcgccgt cgacggccgg
     7681 ccgcggccgc g
//
