LOCUS       sequence-264958-        6333 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 6333)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6333)
  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-314    N-Tag IFP1.4[1] L10 {Esp3I(2x)} E...
FEATURES             Location/Qualifiers
     source          1..6333
                     /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(1492..2066,2067..2069,2070..2078)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1659..1676)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1886..1905
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2079..2081,2082..2084,2085..2798)
                     /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(2124..2145)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2844..2976
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2985..3573
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3038..3058)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3456..3467)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3600..3617)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3606..3717
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3813..3833)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3813..3831)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3852..3868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3852..3868)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3865..3887)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3970..3989)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4035..4090
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4089..4108)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4469..5057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4549..4568)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5228..6019,6020..6088))
                     /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     5851..5870
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6089..6193)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6261..6279
                     /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 caccatggct cgggatccat tgcctttctt cccacccttg tacctcggtg gaccagagat
     1081 caccaccgag aactgcgaga gggaaccgat acacattccc gggtccattc agccccatgg
     1141 tgcgctgctg actgctgacg gacattcagg ggaagtgctg caagtcagcc tgaatgcagc
     1201 aacctttctg ggccaagaac ccactgtgct tcgcgggcag accttggctg ctctgctgcc
     1261 tgagcagtgg ccagccctcc aagcagctct tccacccggt tgtccagacg ccctccagta
     1321 tcgtgccact ctggactggc ctgctgccgg ccaccttagc ctgacagtgc acagggtagc
     1381 tgagctgctc atcctcgaat tcgagcccac cgaggcatgg gggagctctg ccagcggtgg
     1441 cagcacctcc cgagacgccg aggcacgccg tctcctaaga attcagatcc gcccctctcc
     1501 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     1561 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     1621 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     1681 tctgttgaat gtcgtgaagg aagcagttcc tctggtagct tcttgaagac aaacaacgtc
     1741 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     1801 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     1861 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     1921 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt acacatgctt
     1981 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     2041 tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag ggcgaggagc
     2101 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2161 tcagcgtgag gggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
     2221 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgagccacg
     2281 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2341 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcacctaca
     2401 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2461 gcgtcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca
     2521 gccacaacat ctatatcatg gccgtcaagc agaagaacgg catcaaggtg aacttcaaga
     2581 tccgccacaa cgtggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2641 ccatcggcga cggccccgtg ctgctgcccg acagccacta cctgagcacc cagtccgtgc
     2701 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc cgcaccgccg
     2761 ccgggatcac tctcggcatg gacgagctgt acaagtaaac cggtgctcga ggtaccgatg
     2821 aatagctaag gtagaggccg caggtaagta tcaaggttac aagacaggtt taaggagacc
     2881 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
     2941 gtcttactga catccacttt gcctttctct ccacaggtgt cgacaatcaa cctctggatt
     3001 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3061 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3121 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3181 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3241 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3301 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3361 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     3421 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3481 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3541 cgagtcggat ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacgatcagc
     3601 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3661 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3721 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3781 ggattgggaa gacaatagcc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     3841 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctaat tcactcctca
     3901 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     3961 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     4021 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     4081 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     4141 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     4201 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     4261 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     4321 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     4381 cccagtcata gctgtccctc ttctcttatg gagatcggcc ggccgccggc gtcgacgcgc
     4441 gcggccgcgg cgcgccgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4501 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4561 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4621 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4681 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4741 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4801 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4861 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4921 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4981 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5041 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5101 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5161 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5221 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5281 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5341 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5401 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5461 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5521 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5581 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5641 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5701 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5761 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5821 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5881 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5941 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6001 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6061 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6121 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6181 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6241 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtcta cgcgcgcgcc
     6301 ggcggcgcgc cgtcgacggc cggccgcggc cgc
//
