LOCUS       sequence-249206-        6312 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 6312)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6312)
  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-325    C-Tag miRFP670[1] L10 {Esp3I(2x)}...
FEATURES             Location/Qualifiers
     source          1..6312
                     /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(1471..2045,2046..2048,2049..2057)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1638..1655)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1865..1884
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2058..2060,2061..2063,2064..2777)
                     /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(2103..2124)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2823..2955
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2964..3552
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3017..3037)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3435..3446)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3579..3596)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3585..3696
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3792..3812)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3792..3810)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3831..3847)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3831..3847)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3844..3866)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3949..3968)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4014..4069
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4068..4087)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4448..5036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4528..4547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5207..5998,5999..6067))
                     /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     5830..5849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6068..6172)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6240..6258
                     /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 tggctgggca tgctagcggt tctcctgcct ttggcaccgc tagtcactcc aactgcgagc
     1141 acgaggagat ccatcttgct gggagtatcc agcctcacgg agcccttctg gtcgttagcg
     1201 aacacgatca ccgtgtgatt caggcaagcg cgaatgcagc cgagttcctc aacctgggca
     1261 gtgtgctggg agttcccttg gccgaaatcg acggagatct gctgatcaag attctgcccc
     1321 atctcgatcc gacagctgaa ggcatgccag tagcagttag gtgtcgcatt gggaatccgt
     1381 ccaccgagta ttgcggtctg atgcataggc caccagaagg agggctgatt atcgagctgg
     1441 aaagagcggg accctaagaa ttcagatccg cccctctccc tccccccccc ctaacgttac
     1501 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     1561 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     1621 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     1681 agcagttcct ctggtagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     1741 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     1801 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     1861 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     1921 ttgtatggga tctgatctgg ggcctcggta cacatgcttt acatgtgttt agtcgaggtt
     1981 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     2041 ataatatggc cacaaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2101 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgagg ggcgagggcg
     2161 agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2221 ccgtgccctg gcccaccctc gtgaccaccc tgagccacgg cgtgcagtgc ttcgcccgct
     2281 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2341 aggagcgcac catcttcttc aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt
     2401 tcgagggcga caccctggtg aaccgcatcg agctgaaggg cgtcgacttc aaggaggacg
     2461 gcaacatcct ggggcacaag ctggagtaca acttcaacag ccacaacatc tatatcatgg
     2521 ccgtcaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac gtggaggacg
     2581 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2641 tgctgcccga cagccactac ctgagcaccc agtccgtgct gagcaaagac cccaacgaga
     2701 agcgcgatca catggtcctg ctggagttcc gcaccgccgc cgggatcact ctcggcatgg
     2761 acgagctgta caagtaaacc ggtgctcgag gtaccgatga atagctaagg tcgaggccgc
     2821 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga
     2881 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg
     2941 cctttctctc cacaggtgtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     3001 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3061 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3121 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3181 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3241 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3301 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     3361 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3421 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3481 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3541 ccgcctcccc gcctggaatt cgagctcggt acgatcagcc tcgactgtgc cttctagttg
     3601 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3661 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3721 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagccc
     3781 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     3841 tttcctgtgt gaaattgtta tccgctaatt cactcctcag gtgcaggctg cctatcagaa
     3901 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc
     3961 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     4021 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     4081 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     4141 atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat
     4201 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     4261 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     4321 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     4381 tctcttatgg agatcggccg gccgccggcg tcgacgcgcg cggccgcggc gcgccgttgc
     4441 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4501 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4561 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4621 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4681 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4741 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4801 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4861 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4921 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4981 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5581 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5821 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6061 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6121 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6181 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6241 ataggcgtat cacgaggccc tttcgtctac gcgcgcgccg gcggcgcgcc gtcgacggcc
     6301 ggccgcggcc gc
//
