LOCUS       sequence-249427-        7127 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 7127)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7127)
  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-522    N-Tag GFP[2] L32 Tln1(rod) EBPF2
FEATURES             Location/Qualifiers
     source          1..7127
                     /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(2286..2860,2861..2863,2864..2872)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2453..2470)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2680..2699
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2873..2875,2876..2878,2879..3592)
                     /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(2918..2939)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3638..3770
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3779..4367
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3832..3852)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4250..4261)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4394..4411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4400..4511
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4607..4627)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4607..4625)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4646..4662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4646..4662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4659..4681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4764..4783)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4829..4884
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4883..4902)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5263..5851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5343..5362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6022..6813,6814..6882))
                     /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     6645..6664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6883..6987)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7055..7073
                     /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 gacgtgggtg gcggagggag cgaaggcggc ggatccggag ggcctggctc
     1141 tggcggagag gggagcgctg gaggcgggtc agccggtggt gggagtgctg tgtctggtat
     1201 cattgctgac ctcgacacca ccatcatgtt cgctactgct ggcacactta accgtgaggg
     1261 tgctgaaact tttgctgacc accgggaggg tatcttaaag acagcaaagg ttcttgtgga
     1321 ggacaccaag gtcctagtgc agaatgcagc tgggagccag gagaagttgg cacaagccgc
     1381 ccagtcctcc gtggccacca ttacccgcct cgctgatgtg gtcaagctcg gtgcagccag
     1441 cctaggagcc gaagaccctg aaactcaggt ggtgctgatc aatgcagtaa aggacgtagc
     1501 caaggccctg ggtgacctca tcagcgctac gaaggctgca gcgggcaaag ttggggatga
     1561 ccctgcagtg tggcagctca agaactctgc caaggtgatg gtgaccaatg tgacatcatt
     1621 gctcaagaca gtgaaggctg tggaagatga ggccaccaaa ggcacacggg ccctagaggc
     1681 aaccacagag cacatacgtc aggaactggc ggtcttctgt tccccagagc cacctgccaa
     1741 gacctctacc cctgaagact tcatccgaat gaccaagggt attactatgg caacagccaa
     1801 agccgttgct gctggcaatt cctgtcgaca ggaagatgtc attgccacag ccaatctgag
     1861 ccgacgggct attgcggaca tgcttcgggc ttgcaaggaa gcagctttcc acccagaagt
     1921 ggcgcctgat gtacggctcc gagccctgca ctatggccgg gagtgtgcca atggttacct
     1981 ggaactgctg gaccacgtgc tgctgaccct tcagaagcca aacccagacc tgaagcagca
     2041 gctaactgga cactcaaagc gggttgctgg ctctgtgact gagctcatcc aggctgctga
     2101 agccatgaaa ggaacagagt gggtggaccc agaggaccct actgtcattg ctgagaatga
     2161 gctcttggga gccgcagccg ccatcgaggc tgcagccaag aagctggagc agctgaagcc
     2221 cagggccaaa cccaaggagg cggatgagtc cttgaacttt gaggaacaat aagaattcag
     2281 atccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     2341 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     2401 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     2461 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctggt agcttcttga
     2521 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     2581 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     2641 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     2701 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     2761 cggtacacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     2821 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag
     2881 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2941 aaacggccac aagttcagcg tgaggggcga gggcgagggc gatgccacca acggcaagct
     3001 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3061 caccctgagc cacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     3121 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3181 cgacggcacc tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3241 catcgagctg aagggcgtcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3301 gtacaacttc aacagccaca acatctatat catggccgtc aagcagaaga acggcatcaa
     3361 ggtgaacttc aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta
     3421 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacagcc actacctgag
     3481 cacccagtcc gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3541 gttccgcacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaaccggtgc
     3601 tcgaggtacc gatgaatagc taaggtcgag gccgcaggta agtatcaagg ttacaagaca
     3661 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
     3721 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtcgacaa
     3781 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3841 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3901 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3961 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4021 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4081 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4141 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     4201 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4261 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4321 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc
     4381 tcggtacgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4441 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4501 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4561 gacagcaagg gggaggattg ggaagacaat agcccagctt ttgttccctt tagtgagggt
     4621 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4681 taattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     4741 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     4801 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     4861 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     4921 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     4981 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     5041 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     5101 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     5161 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc ggccggccgc
     5221 cggcgtcgac gcgcgcggcc gcggcgcgcc gttgctggcg tttttccata ggctccgccc
     5281 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5341 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5401 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5461 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5521 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5581 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5641 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5701 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5761 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5821 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5881 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5941 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6001 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6061 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6121 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6181 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6241 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6301 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6361 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6421 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6481 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6541 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6601 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6661 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6721 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6781 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6841 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6901 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6961 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     7021 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     7081 tctacgcgcg cgccggcggc gcgccgtcga cggccggccg cggccgc
//
