LOCUS       sequence-269474-        6360 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 6360)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6360)
  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-315    C-Tag IFP1.4[1] L20 {Esp3I(2x)} E...
FEATURES             Location/Qualifiers
     source          1..6360
                     /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(1518..2092,2093..2095,2096..2104)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1685..1702)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1912..1931
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2105..2107,2108..2110,2111..2824)
                     /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(2150..2171)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2870..3002
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3011..3599
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3064..3084)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3482..3493)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3626..3643)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3632..3743
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3839..3859)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3839..3857)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3878..3894)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3878..3894)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3891..3913)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3996..4015)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4061..4116
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4115..4134)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4495..5083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4575..4594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5254..6045,6046..6114))
                     /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     5877..5896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6115..6219)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6287..6305
                     /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 gcaccagtgc tcgggatcca ttgcctttct tcccaccctt
     1141 gtacctcggt ggaccagaga tcaccaccga gaactgcgag agggaaccga tacacattcc
     1201 cgggtccatt cagccccatg gtgcgctgct gactgctgac ggacattcag gggaagtgct
     1261 gcaagtcagc ctgaatgcag caacctttct gggccaagaa cccactgtgc ttcgcgggca
     1321 gaccttggct gctctgctgc ctgagcagtg gccagccctc caagcagctc ttccacccgg
     1381 ttgtccagac gccctccagt atcgtgccac tctggactgg cctgctgccg gccaccttag
     1441 cctgacagtg cacagggtag ctgagctgct catcctcgaa ttcgagccca ccgaggcatg
     1501 gtaagaattc agatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     1561 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     1621 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     1681 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     1741 gtagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     1801 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     1861 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     1921 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     1981 gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     2041 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     2101 aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2161 ggacggcgac gtaaacggcc acaagttcag cgtgaggggc gagggcgagg gcgatgccac
     2221 caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2281 caccctcgtg accaccctga gccacggcgt gcagtgcttc gcccgctacc ccgaccacat
     2341 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2401 cttcttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac
     2461 cctggtgaac cgcatcgagc tgaagggcgt cgacttcaag gaggacggca acatcctggg
     2521 gcacaagctg gagtacaact tcaacagcca caacatctat atcatggccg tcaagcagaa
     2581 gaacggcatc aaggtgaact tcaagatccg ccacaacgtg gaggacggca gcgtgcagct
     2641 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacag
     2701 ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat
     2761 ggtcctgctg gagttccgca ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2821 gtaaaccggt gctcgaggta ccgatgaata gctaaggtcg aggccgcagg taagtatcaa
     2881 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     2941 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     3001 aggtgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3061 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3121 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3181 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3241 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3301 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3361 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3421 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3481 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3541 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3601 tggaattcga gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3661 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3721 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3781 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcccagc ttttgttccc
     3841 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3901 attgttatcc gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     3961 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     4021 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     4081 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     4141 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     4201 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     4261 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     4321 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     4381 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     4441 tcggccggcc gccggcgtcg acgcgcgcgg ccgcggcgcg ccgttgctgg cgtttttcca
     4501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4681 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4921 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6241 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6301 gaggcccttt cgtctacgcg cgcgccggcg gcgcgccgtc gacggccggc cgcggccgcg
//
