LOCUS       sequence-264922-        6330 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 6330)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6330)
  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-313    C-Tag IFP1.4[1] L10 {Esp3I(2x)} E...
FEATURES             Location/Qualifiers
     source          1..6330
                     /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"
     misc_feature    join(1489..2063,2064..2066,2067..2075)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1656..1673)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1883..1902
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2076..2078,2079..2081,2082..2795)
                     /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(2121..2142)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2841..2973
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2982..3570
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3035..3055)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3453..3464)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3597..3614)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3603..3714
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3810..3830)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3810..3828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3849..3865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3849..3865)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3862..3884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3967..3986)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4032..4087
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4086..4105)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4466..5054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4546..4565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5225..6016,6017..6085))
                     /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     5848..5867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6086..6190)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6258..6276
                     /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 cacccgagac gccgaggcac gccgtctccg ggagctctgc cagcggtggc agcaccagtg
     1081 ctcgggatcc attgcctttc ttcccaccct tgtacctcgg tggaccagag atcaccaccg
     1141 agaactgcga gagggaaccg atacacattc ccgggtccat tcagccccat ggtgcgctgc
     1201 tgactgctga cggacattca ggggaagtgc tgcaagtcag cctgaatgca gcaacctttc
     1261 tgggccaaga acccactgtg cttcgcgggc agaccttggc tgctctgctg cctgagcagt
     1321 ggccagccct ccaagcagct cttccacccg gttgtccaga cgccctccag tatcgtgcca
     1381 ctctggactg gcctgctgcc ggccacctta gcctgacagt gcacagggta gctgagctgc
     1441 tcatcctcga attcgagccc accgaggcat ggtaagaatt cagatccgcc cctctccctc
     1501 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     1561 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     1621 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     1681 gttgaatgtc gtgaaggaag cagttcctct ggtagcttct tgaagacaaa caacgtctgt
     1741 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     1801 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     1861 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     1921 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtaca catgctttac
     1981 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     2041 cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc gaggagctgt
     2101 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2161 gcgtgagggg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct
     2221 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg agccacggcg
     2281 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2341 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc acctacaaga
     2401 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggcg
     2461 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc
     2521 acaacatcta tatcatggcc gtcaagcaga agaacggcat caaggtgaac ttcaagatcc
     2581 gccacaacgt ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2641 tcggcgacgg ccccgtgctg ctgcccgaca gccactacct gagcacccag tccgtgctga
     2701 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttccgc accgccgccg
     2761 ggatcactct cggcatggac gagctgtaca agtaaaccgg tgctcgaggt accgatgaat
     2821 agctaaggtc gaggccgcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     2881 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     2941 ttactgacat ccactttgcc tttctctcca caggtgtcga caatcaacct ctggattaca
     3001 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3061 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3121 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3181 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3241 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3301 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3361 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     3421 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3481 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3541 gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac gatcagcctc
     3601 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3661 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3721 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3781 ttgggaagac aatagcccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     3841 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctaattca ctcctcaggt
     3901 gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca
     3961 ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     4021 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     4081 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     4141 ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata
     4201 aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc
     4261 cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc
     4321 cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc
     4381 agtcatagct gtccctcttc tcttatggag atcggccggc cgccggcgtc gacgcgcgcg
     4441 gccgcggcgc gccgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4501 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4561 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4621 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4681 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4741 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4801 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4861 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4921 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccgcca
     4981 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5041 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5101 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5161 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5221 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5281 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5341 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5401 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5461 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5521 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5581 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5641 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5701 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5761 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5821 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5881 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5941 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6001 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6061 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6121 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6181 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     6241 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctacgc gcgcgccggc
     6301 ggcgcgccgt cgacggccgg ccgcggccgc
//
