LOCUS       sequence-249243-        6567 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 6567)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6567)
  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-362    N-Tag Venus210[1] L10 {Esp3I(2x)}...
FEATURES             Location/Qualifiers
     source          1..6567
                     /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(1726..2300,2301..2303,2304..2312)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1893..1910)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2120..2139
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2313..2315,2316..2318,2319..3032)
                     /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(2358..2379)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3078..3210
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3219..3807
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3272..3292)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3690..3701)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3834..3851)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3840..3951
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4047..4067)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4047..4065)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4086..4102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4086..4102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4099..4121)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4204..4223)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4269..4324
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4323..4342)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4703..5291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4783..4802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5462..6253,6254..6322))
                     /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     6085..6104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6323..6427)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6495..6513
                     /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 caccatggtg tcaaagggag aagagttgtt taccggtgtt gttcccatac tggtggaact
     1081 ggatggtgac gtcaatggac acaagtttag cgtttctggc gaaggagaag gcgatgctac
     1141 ctatggcaaa ctgaccctca aactgatctg tacgactgga aagttgcctg taccttggcc
     1201 aacactggtc acaaccttgg gctatgggct gcaatgcttt gcccgttatc ctgatcacat
     1261 gaagcaacat gacttcttta agtctgctat gccagaggga tatgtgcagg aacggactat
     1321 cttcttcaag gacgacggca actacaagac gagagctgag gtgaaatttg agggtgacac
     1381 actggtgaat cgcatagagc tgaaaggcat cgacttcaaa gaagatggga atattctcgg
     1441 gcataaactc gagtacaact acaattccca caacgtgtac atcactgccg ataagcagaa
     1501 gaatgggatt aaggcgaact tcaagatccg acacaacatc gaggatggtg gggtccagct
     1561 tgccgaccat tatcagcaga acacacccat tggggatgga ccagtcctgt tgccggataa
     1621 ccactacctc agttaccaga gcaaactgtc caaagggagc tctgccagcg gtggcagcac
     1681 ctcccgagac gccgaggcac gccgtctcct aagaattcag atccgcccct ctccctcccc
     1741 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     1801 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     1861 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     1921 gaatgtcgtg aaggaagcag ttcctctggt agcttcttga agacaaacaa cgtctgtagc
     1981 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     2041 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     2101 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     2161 ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat gctttacatg
     2221 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     2281 ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca
     2341 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     2401 tgaggggcga gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca
     2461 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgagc cacggcgtgc
     2521 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2581 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcacc tacaagaccc
     2641 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcgtcg
     2701 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca
     2761 acatctatat catggccgtc aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     2821 acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2881 gcgacggccc cgtgctgctg cccgacagcc actacctgag cacccagtcc gtgctgagca
     2941 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttccgcacc gccgccggga
     3001 tcactctcgg catggacgag ctgtacaagt aaaccggtgc tcgaggtacc gatgaatagc
     3061 taaggtagag gccgcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
     3121 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
     3181 ctgacatcca ctttgccttt ctctccacag gtgtcgacaa tcaacctctg gattacaaaa
     3241 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3301 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3361 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3421 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3481 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3541 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3601 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     3661 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3721 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3781 ggatctccct ttgggccgcc tccccgcctg gaattcgagc tcggtacgat cagcctcgac
     3841 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3901 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3961 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4021 ggaagacaat agcccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     4081 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc taattcactc ctcaggtgca
     4141 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     4201 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     4261 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     4321 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     4381 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     4441 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     4501 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     4561 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     4621 catagctgtc cctcttctct tatggagatc ggccggccgc cggcgtcgac gcgcgcggcc
     4681 gcggcgcgcc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4741 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4801 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4861 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4921 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4981 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5041 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5101 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5161 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5221 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5281 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5341 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5401 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5461 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5521 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5581 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5641 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5701 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5761 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5821 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5881 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5941 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6001 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6061 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6121 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6181 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6241 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6301 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6361 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6421 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6481 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctacgcgcg cgccggcggc
     6541 gcgccgtcga cggccggccg cggccgc
//
