LOCUS       sequence-372354-        5516 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 5516)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5516)
  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-399 FLAG miRFP670[1] Stuffer/InteinA
            miR...
FEATURES             Location/Qualifiers
     source          1..5516
                     /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             2156..2179
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2304..2425
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2341..2360)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2395..2414
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2432..2887)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2569..2590)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2781..2800
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2914..3018
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3020..3377
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3020..3211
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3043..3063)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3228..3363
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3412..4206
                     /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             3412..4206
                     /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(3466..3485)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4076..4095
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4394..4413)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4438..4485
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4814..5402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5303..5322
                     /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 gtgctttgcc agagatacgg aagtctacta
     1081 tgagaacgat accgtacctc acatggaatc catcgaagag atgtactcca agtatgccag
     1141 catgaatggc gagttgccat tcgacaatgg ctatgctgtt cctctggaca acgtgtttgt
     1201 ctacactctc gacattgcgt ctggtgagat caagaaaaca cgtgcctcct acatttatcg
     1261 ggaaaaggtc gagaagctga ttgagatcaa gctgagctca gggtatagtc tcaaagtgac
     1321 accgtcccat cccgttctgc tgtttaggga tggactgcag tgggtgccag ctgcagaagt
     1381 gaaacctggt gatgtggttg tcggagtgag agaagaggtg cttcgaaggc gcatcatctc
     1441 aaaaggggaa ctcgaattcc atgaggtgtc tagcgtacgg atcattgact acaacaactg
     1501 ggtgtacgac ttggtcatac ccgaaactca caacttcata gcacccaatg gccttgttct
     1561 gcacaatacc agagcgggac ccagtatcga cctgtctggg acactcgcac cagcactgga
     1621 gcgcataaga actgctggat cactgagggc cctctgtgac gacacagtct tgctctttca
     1681 gcaatgtacg ggctatgatc gagtgatggt gtaccgattc gacgaacagg gtcatggtct
     1741 ggtcttttcc gagtgtcacg tgcctggcct ggagagctac tttggcaacc gctatccctc
     1801 ttctaccgta ccccagatgg ccaggcaact ttacgtccgg caacgggtca gagtgctggt
     1861 ggatgtgacc taccagcctg tgccactcga acctcggctg agcccactca ctggacggga
     1921 tctggacatg agcggctgct tcctgaggtc catgtcacct tgccatctgc agttcttgaa
     1981 ggacatgggc gttcgagcca cactcgcagt ctcactggtt gtaggcggga aactgtgggg
     2041 tcttgtggtg tgtcaccact accttcccag gttcatccgg ttcgagttga gagccatctg
     2101 caaacgtttg gccgagagaa tagccacccg cattactgct ctggaatcca ctggtgatta
     2161 caaggatgac gatgacaagt aagcggccgc gactctagat cataatcagc cataccacat
     2221 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2281 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2341 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2401 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     2461 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2521 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2581 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2641 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2701 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2761 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2821 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2881 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2941 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3001 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3061 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3121 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3181 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3241 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3301 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3361 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3421 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3481 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3541 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3601 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3661 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3721 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3781 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3841 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3901 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     3961 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4021 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4081 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4141 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4201 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4261 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4321 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4381 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4441 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4501 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4561 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4621 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4681 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4741 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4801 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4861 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4921 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4981 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5041 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5101 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5161 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5221 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5281 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5341 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5401 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5461 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
