LOCUS       sequence-249211-        6381 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 6381)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6381)
  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-330    N-Tag miRFP670[1] L32 {Esp3I(2x)}...
FEATURES             Location/Qualifiers
     source          1..6381
                     /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"
     regulatory      1023..1032
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(1540..2114,2115..2117,2118..2126)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1707..1724)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1934..1953
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2127..2129,2130..2132,2133..2846)
                     /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(2172..2193)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          2892..3024
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3033..3621
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3086..3106)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3504..3515)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3648..3665)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3654..3765
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3861..3881)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3861..3879)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3900..3916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3900..3916)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3913..3935)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4018..4037)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4083..4138
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4137..4156)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4517..5105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4597..4616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5276..6067,6068..6136))
                     /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     5899..5918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6137..6241)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6309..6327
                     /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 gctgggcatg ctagcggttc tcctgccttt ggcaccgcta gtcactccaa
     1081 ctgcgagcac gaggagatcc atcttgctgg gagtatccag cctcacggag cccttctggt
     1141 cgttagcgaa cacgatcacc gtgtgattca ggcaagcgcg aatgcagccg agttcctcaa
     1201 cctgggcagt gtgctgggag ttcccttggc cgaaatcgac ggagatctgc tgatcaagat
     1261 tctgccccat ctcgatccga cagctgaagg catgccagta gcagttaggt gtcgcattgg
     1321 gaatccgtcc accgagtatt gcggtctgat gcataggcca ccagaaggag ggctgattat
     1381 cgagctggaa agagcgggac ccgacgtggg tggcggaggg agcgaaggcg gcggatccgg
     1441 agggcctggc tctggcggag aggggagcgc tggaggcggg tcagccggtg gtgggagtcg
     1501 agacgccgag gcacgccgtc tcctaagaat tcagatccgc ccctctccct cccccccccc
     1561 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     1621 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     1681 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     1741 cgtgaaggaa gcagttcctc tggtagcttc ttgaagacaa acaacgtctg tagcgaccct
     1801 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     1861 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     1921 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     1981 ggtaccccat tgtatgggat ctgatctggg gcctcggtac acatgcttta catgtgttta
     2041 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     2101 aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     2161 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgaggg
     2221 gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg
     2281 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gagccacggc gtgcagtgct
     2341 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2401 gctacgtcca ggagcgcacc atcttcttca aggacgacgg cacctacaag acccgcgccg
     2461 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc gtcgacttca
     2521 aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc cacaacatct
     2581 atatcatggc cgtcaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaacg
     2641 tggaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2701 gccccgtgct gctgcccgac agccactacc tgagcaccca gtccgtgctg agcaaagacc
     2761 ccaacgagaa gcgcgatcac atggtcctgc tggagttccg caccgccgcc gggatcactc
     2821 tcggcatgga cgagctgtac aagtaaaccg gtgctcgagg taccgatgaa tagctaaggt
     2881 cgaggccgca ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg
     2941 gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca
     3001 tccactttgc ctttctctcc acaggtgtcg acaatcaacc tctggattac aaaatttgtg
     3061 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3121 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3181 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3241 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3301 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3361 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3421 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3481 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3541 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3601 ccctttgggc cgcctccccg cctggaattc gagctcggta cgatcagcct cgactgtgcc
     3661 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3721 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3781 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3841 caatagccca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     3901 tcatagctgt ttcctgtgtg aaattgttat ccgctaattc actcctcagg tgcaggctgc
     3961 ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct
     4021 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     4081 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     4141 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     4201 tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca
     4261 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     4321 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     4381 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     4441 tgtccctctt ctcttatgga gatcggccgg ccgccggcgt cgacgcgcgc ggccgcggcg
     4501 cgccgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4561 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4621 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4681 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4741 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4801 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4861 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4921 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4981 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5041 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5101 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5161 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5221 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5281 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5341 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5401 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5461 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5521 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5581 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5641 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5701 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5761 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5821 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5881 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5941 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6001 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6061 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6121 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6181 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6241 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     6301 cctataaaaa taggcgtatc acgaggccct ttcgtctacg cgcgcgccgg cggcgcgccg
     6361 tcgacggccg gccgcggccg c
//
