LOCUS       sequence-249533-        5411 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 5411)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5411)
  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-407 FLAG miRFP670[1] Stuffer/InteinC
            miR...
FEATURES             Location/Qualifiers
     source          1..5411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             644..667
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2051..2074
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2199..2320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2236..2255)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2290..2309
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2327..2782)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2464..2485)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2676..2695
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2809..2913
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2915..3272
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2915..3106
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2938..2958)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3123..3258
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3307..4101
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3307..4101
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3361..3380)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3971..3990
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4289..4308)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4333..4380
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4709..5297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5198..5217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcgccacc atggactaca aagacgatga
      661 cgataaagga agtgtggctg ggcatgctag cggttctcct gcctttggca ccgctagtca
      721 ctccaactgc gagcacgagg agatccatct tgctgggagt atccagcctc acggagccct
      781 tctggtcgtt agcgaacacg atcaccgtgt gattcaggca agcgcgaatg cagccgagtt
      841 cctcaacctg ggcagtgtgc tgggagttcc cttggccgaa atcgacggag atctgctgat
      901 caagattctg ccccatctcg atccgacagc tgaaggcatg ccagtagcag ttaggtgtcg
      961 cattgggaat ccgtccaccg agtattgcgg tctgatgcat aggccaccag aaggagggct
     1021 gattatcgag ctggaaagag cgggaccctc ttgtctgtcc tatgagactg agatcctgac
     1081 agttgagtat ggtctgcttc ccataggcaa gatcgtggag aaaaggatcg aatgtaccgt
     1141 gtactcagtc gacaataacg gcaacatcta cactcaacct gttgcacagt ggcatgatag
     1201 aggagaacag gaagtgttcg agtactgctt ggaggatggg agtctgatac gagccacgaa
     1261 agaccacaag ttcatgaccg tagatggaca gatgctgcca attgacgaga tctttgagcg
     1321 ggaattggac ctgatgaggg tggataatct tccgaacatc aagattgcta cacgcaagta
     1381 tctcgggaaa cagaacgtct acgacattgg tgtggaacgt gaccacaact ttgcgctcaa
     1441 gaatggcttc attgccagca attgcagagc gggacccagt atcgacctgt ctgggacact
     1501 cgcaccagca ctggagcgca taagaactgc tggatcactg agggccctct gtgacgacac
     1561 agtcttgctc tttcagcaat gtacgggcta tgatcgagtg atggtgtacc gattcgacga
     1621 acagggtcat ggtctggtct tttccgagtg tcacgtgcct ggcctggaga gctactttgg
     1681 caaccgctat ccctcttcta ccgtacccca gatggccagg caactttacg tccggcaacg
     1741 ggtcagagtg ctggtggatg tgacctacca gcctgtgcca ctcgaacctc ggctgagccc
     1801 actcactgga cgggatctgg acatgagcgg ctgcttcctg aggtccatgt caccttgcca
     1861 tctgcagttc ttgaaggaca tgggcgttcg agccacactc gcagtctcac tggttgtagg
     1921 cgggaaactg tggggtcttg tggtgtgtca ccactacctt cccaggttca tccggttcga
     1981 gttgagagcc atctgcaaac gtttggccga gagaatagcc acccgcatta ctgctctgga
     2041 atccactggt gattacaagg atgacgatga caagtaagcg gccgcgactc tagatcataa
     2101 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2161 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2281 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     2341 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2401 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2461 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2521 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2581 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2641 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2701 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2761 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2821 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2881 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2941 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3001 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3121 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3181 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3241 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3301 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3361 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3421 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3481 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3541 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3601 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3661 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3721 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3781 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3841 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3901 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3961 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4021 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4081 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4141 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4201 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4261 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4321 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4381 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4441 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4501 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4561 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4621 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4681 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4741 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4801 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4861 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4921 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4981 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5041 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5101 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5161 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5221 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5281 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5341 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5401 ccgccatgca t
//
