LOCUS       sequence-249508-        5600 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 5600)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 5600)
  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-020    FLAG miRFP670[1] IRES miRFP670[2]
FEATURES             Location/Qualifiers
     source          1..5600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        173..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      747..756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             756..779
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(1177..1751,1752..1754,1755..1763)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1344..1361)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1571..1590
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2352..2375
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2500..2621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2537..2556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2591..2610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2628..3083)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2765..2786)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2977..2996
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3110..3214
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3216..3573
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3216..3407
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3239..3259)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3424..3559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3608..4402
                     /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             3608..4402
                     /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(3662..3681)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4272..4291
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4590..4609)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4634..4681
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5010..5598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5499..5518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
       61 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
      721 agatctcgag ctcaagcttc gaattcgcca ccatggacta caaagacgat gacgataaag
      781 gaagtgtggc tgggcatgct agcggttctc ctgcctttgg caccgctagt cactccaact
      841 gcgagcacga ggagatccat cttgctggga gtatccagcc tcacggagcc cttctggtcg
      901 ttagcgaaca cgatcaccgt gtgattcagg caagcgcgaa tgcagccgag ttcctcaacc
      961 tgggcagtgt gctgggagtt cccttggccg aaatcgacgg agatctgctg atcaagattc
     1021 tgccccatct cgatccgaca gctgaaggca tgccagtagc agttaggtgt cgcattggga
     1081 atccgtccac cgagtattgc ggtctgatgc ataggccacc agaaggaggg ctgattatcg
     1141 agctggaaag agcgggaccc taagaattca gatccgcccc tctccctccc ccccccctaa
     1201 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     1261 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     1321 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     1381 gaaggaagca gttcctctgg tagcttcttg aagacaaaca acgtctgtag cgaccctttg
     1441 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     1501 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     1561 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     1621 accccattgt atgggatctg atctggggcc tcggtacaca tgctttacat gtgtttagtc
     1681 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     1741 acgatgataa tatggccaca accatgagag cgggacccag tatcgacctg tctgggacac
     1801 tcgcaccagc actggagcgc ataagaactg ctggatcact gagggccctc tgtgacgaca
     1861 cagtcttgct ctttcagcaa tgtacgggct atgatcgagt gatggtgtac cgattcgacg
     1921 aacagggtca tggtctggtc ttttccgagt gtcacgtgcc tggcctggag agctactttg
     1981 gcaaccgcta tccctcttct accgtacccc agatggccag gcaactttac gtccggcaac
     2041 gggtcagagt gctggtggat gtgacctacc agcctgtgcc actcgaacct cggctgagcc
     2101 cactcactgg acgggatctg gacatgagcg gctgcttcct gaggtccatg tcaccttgcc
     2161 atctgcagtt cttgaaggac atgggcgttc gagccacact cgcagtctca ctggttgtag
     2221 gcgggaaact gtggggtctt gtggtgtgtc accactacct tcccaggttc atccggttcg
     2281 agttgagagc catctgcaaa cgtttggccg agagaatagc cacccgcatt actgctctgg
     2341 aatccactgg tgattacaag gatgacgatg acaagtaagc ggccgcgact ctagatcata
     2401 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2461 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2521 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2581 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2641 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2701 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2761 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2821 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2881 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2941 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3001 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3061 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3181 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3241 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3301 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3361 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3421 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3481 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3541 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3601 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3661 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3721 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3781 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3841 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3901 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3961 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4021 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4081 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4141 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4201 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4261 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4321 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4381 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4441 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4501 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4561 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4621 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4681 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4741 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4801 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4861 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4921 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4981 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5041 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5101 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     5161 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5221 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5281 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5341 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5401 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5461 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5521 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5581 gggcggagcc tatggaaaaa
//
