LOCUS       sequence-321655-        5568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of RanQ69L with tag-BFP; contains miniAux 
            domain for degredation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5568)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 5568)
  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: BFP-RanQ69L
FEATURES             Location/Qualifiers
     source          1..5568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        55..358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        359..562
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     513..533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             598..801
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             817..1509
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKL"
     polyA_signal    2350..2471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2387..2406)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2441..2460
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2478..2933)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2615..2636)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2827..2846
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2960..3064
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3066..3423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3066..3257
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3089..3109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3274..3409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3458..4252
                     /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             3458..4252
                     /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(3512..3531)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4122..4141
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4440..4459)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4484..4531
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4860..5448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5349..5368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
       61 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      121 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      181 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      241 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      301 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      361 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      421 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      481 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      541 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc caccatgaag
      601 gagaagagtg cttgtcctaa agatccagcc aaacctccgg ccaaggcaca agttgtggga
      661 tggccaccgg tgagatcata ccggaagaac gtgatggttt cctgccaaaa atcaagcggt
      721 ggcccggagg cggcggcgtt cgtgaaggta tcaatggacg gagcaccgta cttgaggaaa
      781 atcgatttga ggatgtataa aggtggcgga ggctccagcg agctgattaa ggagaacatg
      841 cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag
      901 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc
      961 cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc
     1021 atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca
     1081 tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     1141 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac
     1201 ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc
     1261 gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat
     1321 ctgatcgcaa acatcaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg
     1381 cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc
     1441 tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg
     1501 cacaagcttg gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg
     1561 aattctggaa tggctgcgca gggagagccc caggtccagt tcaaacttgt attggttggt
     1621 gatggtggta ctggaaaaac gaccttcgtg aaacgtcatt tgactggtga atttgagaag
     1681 aagtatgtag ccaccttggg tgttgaggtt catcccctag tgttccacac caacagagga
     1741 cctattaagt tcaatgtatg ggacacagcc ggcctggaga aattcggtgg actgagagat
     1801 ggctattata tccaagccca gtgtgccatc ataatgtttg atgtaacatc gagagttact
     1861 tacaagaatg tgcctaactg gcatagagat ctggtacgag tgtgtgaaaa catccccatt
     1921 gtgttgtgtg gcaacaaagt ggatattaag gacaggaaag tgaaggcgaa atccattgtc
     1981 ttccaccgaa agaagaatct tcagtactac gacatttctg ccaaaagtaa ctacaacttt
     2041 gaaaagccct tcctctggct tgctaggaag ctcattggag accctaactt ggaatttgtt
     2101 gccatgcctg ctctcgcccc accagaagtt gtcatggacc cagctttggc agcacagtat
     2161 gagcacgact tagaggttgc tcagacaact gctctcccgg atgaggatga tgacctgtga
     2221 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     2281 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2341 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2401 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2461 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2521 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2581 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2641 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2701 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2761 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2821 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2881 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     2941 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3001 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3061 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3241 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3361 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3421 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     3661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3961 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     4021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4381 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     4441 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4501 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4561 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4621 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4681 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     4741 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4801 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4861 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4921 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4981 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5041 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5101 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5161 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5221 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5281 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5341 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5401 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5461 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5521 ttatcccctg attctgtgga taaccgtatt accgccatgc attagtta
//
