LOCUS       sequence-249528-        5564 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 5564)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5564)
  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-402 FLAG IFP1.4[1] Stuffer/InteinB IFP1....
FEATURES             Location/Qualifiers
     source          1..5564
                     /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             2204..2227
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2352..2473
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2389..2408)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2443..2462
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2480..2935)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2617..2638)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2829..2848
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2962..3066
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3068..3425
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3068..3259
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3091..3111)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3276..3411
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3460..4254
                     /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             3460..4254
                     /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(3514..3533)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4124..4143
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4442..4461)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4486..4533
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4862..5450
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5351..5370
                     /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 gcgtgtcagg
     1081 agaaactcct gtgtaccttg ctgatggcaa gaccatcaag atcaaggacc tgtattcatc
     1141 cgagaggaag aaggaggata acatcgtgga ggctggatct ggggaagaga tcatccacct
     1201 gaaagacccg attcagatct actcctacgt ggatgggacc atagtacgaa gcagaagcag
     1261 actgctgtac aaaggcaaat ccagctatct ggtgcgtatt gagactatcg gaggtaggtc
     1321 agtcagtgtg acaccagtcc ataagctctt cgtccttacg gagaaaggca tagaggaagt
     1381 catggcctcc aatctgaaag tgggtgacat gattgcggct gtagccgaaa gtgagagcga
     1441 agcacgggat tgtgggatgt ctgaggaatg cgttatggaa gcagaggttt acacctctct
     1501 ggaagccaca tttgatcggg tcaagagcat tgcctacgag aaaggggact ttgacgtgta
     1561 tgacctcagt gtgcccgaat atggtcgcaa cttcattggc ggagaaggct tgttggttct
     1621 ccacaataca gacagcattg gaccccatgc gctgaggaac gctatgttcg cccttgaatc
     1681 tgctcccaat ctccgagcac tggctgaagt cgccacacag acagtcaggg agctgacagg
     1741 ctttgatcgg gtgatgctgt acaagtttgc cccagatgca acgggtgaga tgatcgcaga
     1801 agccagacgg gaagggatgc aggccttcct gggccacaga tttccggcct cacatacgcc
     1861 tgctcaggcc agagcactgt acacccgaca cctcctgaga ctgactgcgg acactagagc
     1921 agctgccgtg ccactcgatc ccgtgctcaa cccacagacc aatgcgccta cccctttggg
     1981 aggagccgta ctgagggcca catcccctat gcacatgcag tatctgcgca acatgggcgt
     2041 tgggagtagt ctgtctgtgt ccgttgtcgt tggagggcaa ctgtggggct tgatcgtctg
     2101 ccaccatcag acaccgtatg tgcttcctcc tgacttgcgt acaactctgg aggagctggg
     2161 ccgcaagctt agcggccagg tgcagcggaa agaggccact ggtgattaca aggatgacga
     2221 tgacaagtaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     2281 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2341 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2401 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2461 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2521 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     2581 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2641 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2701 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2761 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2821 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     2881 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     2941 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3001 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3061 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3241 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3361 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3421 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3481 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3541 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3601 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     3661 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3721 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3781 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3841 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3901 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3961 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     4021 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4081 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4141 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4201 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4261 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4321 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4381 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     4441 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     4501 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4561 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4621 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4681 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     4741 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4801 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4861 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4921 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4981 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5041 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5101 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5161 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5221 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5281 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5341 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5401 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5461 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5521 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
