LOCUS       sequence-249529-        5558 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 5558)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5558)
  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-403 FLAG miRFP670[1] Stuffer/InteinB
            miR...
FEATURES             Location/Qualifiers
     source          1..5558
                     /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             2198..2221
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2346..2467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2383..2402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2437..2456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2611..2632)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2823..2842
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2956..3060
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3062..3419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3062..3253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3085..3105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3270..3405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3454..4248
                     /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             3454..4248
                     /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(3508..3527)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4118..4137
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4436..4455)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4480..4527
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4856..5444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5345..5364
                     /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 cgggacccaa gtgcgtgtca ggagaaactc ctgtgtacct
     1081 tgctgatggc aagaccatca agatcaagga cctgtattca tccgagagga agaaggagga
     1141 taacatcgtg gaggctggat ctggggaaga gatcatccac ctgaaagacc cgattcagat
     1201 ctactcctac gtggatggga ccatagtacg aagcagaagc agactgctgt acaaaggcaa
     1261 atccagctat ctggtgcgta ttgagactat cggaggtagg tcagtcagtg tgacaccagt
     1321 ccataagctc ttcgtcctta cggagaaagg catagaggaa gtcatggcct ccaatctgaa
     1381 agtgggtgac atgattgcgg ctgtagccga aagtgagagc gaagcacggg attgtgggat
     1441 gtctgaggaa tgcgttatgg aagcagaggt ttacacctct ctggaagcca catttgatcg
     1501 ggtcaagagc attgcctacg agaaagggga ctttgacgtg tatgacctca gtgtgcccga
     1561 atatggtcgc aacttcattg gcggagaagg cttgttggtt ctccacaata caagagcggg
     1621 acccagtatc gacctgtctg ggacactcgc accagcactg gagcgcataa gaactgctgg
     1681 atcactgagg gccctctgtg acgacacagt cttgctcttt cagcaatgta cgggctatga
     1741 tcgagtgatg gtgtaccgat tcgacgaaca gggtcatggt ctggtctttt ccgagtgtca
     1801 cgtgcctggc ctggagagct actttggcaa ccgctatccc tcttctaccg taccccagat
     1861 ggccaggcaa ctttacgtcc ggcaacgggt cagagtgctg gtggatgtga cctaccagcc
     1921 tgtgccactc gaacctcggc tgagcccact cactggacgg gatctggaca tgagcggctg
     1981 cttcctgagg tccatgtcac cttgccatct gcagttcttg aaggacatgg gcgttcgagc
     2041 cacactcgca gtctcactgg ttgtaggcgg gaaactgtgg ggtcttgtgg tgtgtcacca
     2101 ctaccttccc aggttcatcc ggttcgagtt gagagccatc tgcaaacgtt tggccgagag
     2161 aatagccacc cgcattactg ctctggaatc cactggtgat tacaaggatg acgatgacaa
     2221 gtaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     2281 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     2341 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2401 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2461 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     2941 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3001 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3061 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     3421 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3481 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3541 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3601 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     3661 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3721 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3781 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3841 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3901 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3961 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     4021 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4081 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4141 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4201 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4261 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4321 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4381 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     4441 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4501 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4561 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4621 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4681 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     4741 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4801 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4861 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4921 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4981 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5041 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5101 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5161 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5221 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5281 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5341 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5401 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5461 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5521 cccctgattc tgtggataac cgtattaccg ccatgcat
//
