LOCUS       sequence-249425-        7091 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 7091)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7091)
  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-520    N-Tag GFP[2] L20 Tln1(rod) EBPF2
FEATURES             Location/Qualifiers
     source          1..7091
                     /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"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(2250..2824,2825..2827,2828..2836)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2417..2434)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2644..2663
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2837..2839,2840..2842,2843..3556)
                     /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(2882..2903)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3602..3734
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3743..4331
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3796..3816)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4214..4225)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4358..4375)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4364..4475
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4571..4591)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4571..4589)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4610..4626)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4610..4626)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4623..4645)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4728..4747)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4793..4848
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4847..4866)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5227..5815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5307..5326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5986..6777,6778..6846))
                     /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     6609..6628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6847..6951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7019..7037
                     /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 caccatggaa aagagggacc acatggtgct gctcgagtac gtcacagcag ctggcatcac
     1081 cgatgccagc gggagtcctg gcggtggatc cggaggctct gggagctctg ccagcggtgg
     1141 cagcacctcc gctgtgtctg gtatcattgc tgacctcgac accaccatca tgttcgctac
     1201 tgctggcaca cttaaccgtg agggtgctga aacttttgct gaccaccggg agggtatctt
     1261 aaagacagca aaggttcttg tggaggacac caaggtccta gtgcagaatg cagctgggag
     1321 ccaggagaag ttggcacaag ccgcccagtc ctccgtggcc accattaccc gcctcgctga
     1381 tgtggtcaag ctcggtgcag ccagcctagg agccgaagac cctgaaactc aggtggtgct
     1441 gatcaatgca gtaaaggacg tagccaaggc cctgggtgac ctcatcagcg ctacgaaggc
     1501 tgcagcgggc aaagttgggg atgaccctgc agtgtggcag ctcaagaact ctgccaaggt
     1561 gatggtgacc aatgtgacat cattgctcaa gacagtgaag gctgtggaag atgaggccac
     1621 caaaggcaca cgggccctag aggcaaccac agagcacata cgtcaggaac tggcggtctt
     1681 ctgttcccca gagccacctg ccaagacctc tacccctgaa gacttcatcc gaatgaccaa
     1741 gggtattact atggcaacag ccaaagccgt tgctgctggc aattcctgtc gacaggaaga
     1801 tgtcattgcc acagccaatc tgagccgacg ggctattgcg gacatgcttc gggcttgcaa
     1861 ggaagcagct ttccacccag aagtggcgcc tgatgtacgg ctccgagccc tgcactatgg
     1921 ccgggagtgt gccaatggtt acctggaact gctggaccac gtgctgctga cccttcagaa
     1981 gccaaaccca gacctgaagc agcagctaac tggacactca aagcgggttg ctggctctgt
     2041 gactgagctc atccaggctg ctgaagccat gaaaggaaca gagtgggtgg acccagagga
     2101 ccctactgtc attgctgaga atgagctctt gggagccgca gccgccatcg aggctgcagc
     2161 caagaagctg gagcagctga agcccagggc caaacccaag gaggcggatg agtccttgaa
     2221 ctttgaggaa caataagaat tcagatccgc ccctctccct cccccccccc taacgttact
     2281 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     2341 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     2401 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     2461 gcagttcctc tggtagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     2521 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     2581 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     2641 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     2701 tgtatgggat ctgatctggg gcctcggtac acatgcttta catgtgttta gtcgaggtta
     2761 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     2821 taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2881 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgaggg gcgagggcga
     2941 gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3001 cgtgccctgg cccaccctcg tgaccaccct gagccacggc gtgcagtgct tcgcccgcta
     3061 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3121 ggagcgcacc atcttcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt
     3181 cgagggcgac accctggtga accgcatcga gctgaagggc gtcgacttca aggaggacgg
     3241 caacatcctg gggcacaagc tggagtacaa cttcaacagc cacaacatct atatcatggc
     3301 cgtcaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaacg tggaggacgg
     3361 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3421 gctgcccgac agccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa
     3481 gcgcgatcac atggtcctgc tggagttccg caccgccgcc gggatcactc tcggcatgga
     3541 cgagctgtac aagtaaaccg gtgctcgagg taccgatgaa tagctaaggt agaggccgca
     3601 ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag
     3661 acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc
     3721 ctttctctcc acaggtgtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3781 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3841 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3901 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3961 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4021 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4081 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     4141 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4201 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4261 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4321 cgcctccccg cctggaattc gagctcggta cgatcagcct cgactgtgcc ttctagttgc
     4381 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     4441 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     4501 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagccca
     4561 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     4621 ttcctgtgtg aaattgttat ccgctaattc actcctcagg tgcaggctgc ctatcagaag
     4681 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     4741 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     4801 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     4861 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     4921 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     4981 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     5041 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     5101 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     5161 ctcttatgga gatcggccgg ccgccggcgt cgacgcgcgc ggccgcggcg cgccgttgct
     5221 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5281 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5341 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5401 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5461 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5521 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5581 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5641 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5701 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5761 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5821 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5881 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5941 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6001 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6061 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6121 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6181 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6241 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6301 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6361 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6421 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6481 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6541 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6601 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6661 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6721 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6781 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6841 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6901 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6961 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     7021 taggcgtatc acgaggccct ttcgtctacg cgcgcgccgg cggcgcgccg tcgacggccg
     7081 gccgcggccg c
//
