LOCUS       sequence-249208-        6342 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 6342)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6342)
  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-327    C-Tag miRFP670[1] L20 {Esp3I(2x)}...
FEATURES             Location/Qualifiers
     source          1..6342
                     /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(1500..2074,2075..2077,2078..2086)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1667..1684)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1894..1913
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2087..2089,2090..2092,2093..2806)
                     /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(2132..2153)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2852..2984
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2993..3581
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3046..3066)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3464..3475)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3608..3625)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3614..3725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3821..3841)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3821..3839)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3860..3876)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3860..3876)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3873..3895)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3978..3997)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4043..4098
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4097..4116)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4477..5065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4557..4576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5236..6027,6028..6096))
                     /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     5859..5878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6097..6201)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6269..6287
                     /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 ggctgggcat gctagcggtt ctcctgcctt
     1141 tggcaccgct agtcactcca actgcgagca cgaggagatc catcttgctg ggagtatcca
     1201 gcctcacgga gcccttctgg tcgttagcga acacgatcac cgtgtgattc aggcaagcgc
     1261 gaatgcagcc gagttcctca acctgggcag tgtgctggga gttcccttgg ccgaaatcga
     1321 cggagatctg ctgatcaaga ttctgcccca tctcgatccg acagctgaag gcatgccagt
     1381 agcagttagg tgtcgcattg ggaatccgtc caccgagtat tgcggtctga tgcataggcc
     1441 accagaagga gggctgatta tcgagctgga aagagcggga ccctaagaat tcagatccgc
     1501 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     1561 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     1621 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     1681 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggtagcttc ttgaagacaa
     1741 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     1801 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     1861 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     1921 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac
     1981 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     2041 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg
     2101 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2161 ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc accaacggca agctgaccct
     2221 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2281 gagccacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     2341 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2401 cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2461 gctgaagggc gtcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2521 cttcaacagc cacaacatct atatcatggc cgtcaagcag aagaacggca tcaaggtgaa
     2581 cttcaagatc cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc actaccagca
     2641 gaacaccccc atcggcgacg gccccgtgct gctgcccgac agccactacc tgagcaccca
     2701 gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttccg
     2761 caccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaaccg gtgctcgagg
     2821 taccgatgaa tagctaaggt cgaggccgca ggtaagtatc aaggttacaa gacaggttta
     2881 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
     2941 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcg acaatcaacc
     3001 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3061 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3121 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3181 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3241 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3301 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3361 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     3421 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3481 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3541 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaattc gagctcggta
     3601 cgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3661 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3721 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3781 aagggggagg attgggaaga caatagccca gcttttgttc cctttagtga gggttaattg
     3841 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctaattc
     3901 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct
     3961 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc
     4021 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
     4081 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag
     4141 aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa
     4201 ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc
     4261 catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt
     4321 tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc
     4381 tgactactcc cagtcatagc tgtccctctt ctcttatgga gatcggccgg ccgccggcgt
     4441 cgacgcgcgc ggccgcggcg cgccgttgct ggcgtttttc cataggctcc gcccccctga
     4501 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4561 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4621 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4681 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4741 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4801 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4861 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4921 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4981 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5041 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5101 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5161 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5221 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5281 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5341 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5401 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5461 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5521 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5581 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5641 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5701 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5761 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5821 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5881 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5941 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6001 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6061 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6181 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6241 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctacg
     6301 cgcgcgccgg cggcgcgccg tcgacggccg gccgcggccg cg
//
