LOCUS       sequence-249524-        5522 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 5522)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5522)
  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-398 FLAG IFP1.4[1] Stuffer/InteinA IFP1....
FEATURES             Location/Qualifiers
     source          1..5522
                     /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             2162..2185
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2310..2431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2347..2366)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2401..2420
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2438..2893)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2575..2596)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2787..2806
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2920..3024
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3026..3383
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3026..3217
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3049..3069)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3234..3369
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3418..4212
                     /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             3418..4212
                     /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(3472..3491)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4082..4101
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4400..4419)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4444..4491
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4820..5408
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5309..5328
                     /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 agtgctcggg atccattgcc tttcttccca cccttgtacc tcggtggacc
      721 agagatcacc accgagaact gcgagaggga accgatacac attcccgggt ccattcagcc
      781 ccatggtgcg ctgctgactg ctgacggaca ttcaggggaa gtgctgcaag tcagcctgaa
      841 tgcagcaacc tttctgggcc aagaacccac tgtgcttcgc gggcagacct tggctgctct
      901 gctgcctgag cagtggccag ccctccaagc agctcttcca cccggttgtc cagacgccct
      961 ccagtatcgt gccactctgg actggcctgc tgccggccac cttagcctga cagtgcacag
     1021 ggtagctgag ctgctcatcc tcgaattcga gcccaccgag gcatggaagt gctttgccag
     1081 agatacggaa gtctactatg agaacgatac cgtacctcac atggaatcca tcgaagagat
     1141 gtactccaag tatgccagca tgaatggcga gttgccattc gacaatggct atgctgttcc
     1201 tctggacaac gtgtttgtct acactctcga cattgcgtct ggtgagatca agaaaacacg
     1261 tgcctcctac atttatcggg aaaaggtcga gaagctgatt gagatcaagc tgagctcagg
     1321 gtatagtctc aaagtgacac cgtcccatcc cgttctgctg tttagggatg gactgcagtg
     1381 ggtgccagct gcagaagtga aacctggtga tgtggttgtc ggagtgagag aagaggtgct
     1441 tcgaaggcgc atcatctcaa aaggggaact cgaattccat gaggtgtcta gcgtacggat
     1501 cattgactac aacaactggg tgtacgactt ggtcataccc gaaactcaca acttcatagc
     1561 acccaatggc cttgttctgc acaataccga cagcattgga ccccatgcgc tgaggaacgc
     1621 tatgttcgcc cttgaatctg ctcccaatct ccgagcactg gctgaagtcg ccacacagac
     1681 agtcagggag ctgacaggct ttgatcgggt gatgctgtac aagtttgccc cagatgcaac
     1741 gggtgagatg atcgcagaag ccagacggga agggatgcag gccttcctgg gccacagatt
     1801 tccggcctca catacgcctg ctcaggccag agcactgtac acccgacacc tcctgagact
     1861 gactgcggac actagagcag ctgccgtgcc actcgatccc gtgctcaacc cacagaccaa
     1921 tgcgcctacc cctttgggag gagccgtact gagggccaca tcccctatgc acatgcagta
     1981 tctgcgcaac atgggcgttg ggagtagtct gtctgtgtcc gttgtcgttg gagggcaact
     2041 gtggggcttg atcgtctgcc accatcagac accgtatgtg cttcctcctg acttgcgtac
     2101 aactctggag gagctgggcc gcaagcttag cggccaggtg cagcggaaag aggccactgg
     2161 tgattacaag gatgacgatg acaagtaagc ggccgcgact ctagatcata atcagccata
     2221 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2281 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2341 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2401 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     2461 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     2521 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     2581 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     2641 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     2701 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     2761 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     2821 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     2881 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2941 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3001 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3061 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3121 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3181 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3241 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3301 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3361 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3421 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3481 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3541 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     3601 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3661 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3721 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3781 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3841 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3901 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     3961 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4021 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4081 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4141 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4201 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4261 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4321 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4381 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4441 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     4501 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     4561 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     4621 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     4681 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4741 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4801 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4861 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4921 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4981 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5041 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5101 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5161 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5221 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5281 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5341 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5401 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5461 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     5521 at
//
