LOCUS       sequence-215998-        7054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Translocating Tiam1 to iLID and activate Rac1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7054)
  AUTHORS   Van Geel O, Hartsuiker R, Gadella TWJ
  TITLE     Increasing spatial resolution of photoregulated GTPases through 
            immobilized peripheral membrane proteins.
  JOURNAL   Small GTPases. 2018 Sep 5:1-10. doi: 10.1080/21541248.2018.1507411.
  PUBMED    30182785
REFERENCE   2  (bases 1 to 7054)
  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"; journalName: "Small 
            GTPases. 2018 Sep 5:1-10. doi: 10.1080/21541248.2018.1507411."
COMMENT     Sequence Label: 2xsspB-mSc-sspB-Tiam1
FEATURES             Location/Qualifiers
     source          1..7054
                     /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"
     misc_feature    609..665
                     /label=MCS
                     /note="multiple cloning site"
     CDS             741..1070
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein"
                     /label=SspB nano
                     /note="dimerizes with iLID at high affinity to create iLID 
                     nano (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNARFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     CDS             1161..1490
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein"
                     /label=SspB nano
                     /note="dimerizes with iLID at high affinity to create iLID 
                     nano (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNARFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     regulatory      1503..1512
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1509..2204
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1642..1660)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1910..1929
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2277..2606
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein"
                     /label=SspB nano
                     /note="dimerizes with iLID at high affinity to create iLID 
                     nano (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNARFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     polyA_signal    3842..3963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3879..3898)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3933..3952
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3970..4425)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4107..4128)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4319..4338
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4452..4556
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4558..4915
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4558..4749
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4581..4601)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4766..4901
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4950..5744
                     /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             4950..5744
                     /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(5004..5023)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5614..5633
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5932..5951)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5976..6023
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6352..6940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6841..6860
                     /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 attctgcagt cgacggtacc gcgggcccgg
      661 gatccatggg actcagatct cgagctcaag cttcgaacga attcggcatt gatctgagcg
      721 gcctgaccct gcaggaattc agctccccga aacgccctaa gctgctgcgt gaatattacg
      781 attggctggt tgataacagc tttaccccat atctggtggt ggatgccaca tacctgggcg
      841 tgaacgtgcc cgtggagtat gtgaaagacg gtcagatcgt gctgaatctg tctgcaagtg
      901 cgaccggcaa cctgcaactg acaaatgatt ttatccagtt caacgcccgc tttaagggcg
      961 tgtctcgtga actgtatatc ccgatgggtg ccgctctggc catttacgct cgcgagaacg
     1021 gcgatggtgt gatgttcgaa ccagaagaaa tctatgacga gctgaatatt ggtccaccgg
     1081 gatccatggg actcagatct cgagctcaag cttcgaacga attcggcatt gatctgagcg
     1141 gcctgaccct gcaggaattc agctccccga aacgccctaa gctgctgcgt gaatattacg
     1201 attggctggt tgataacagc tttaccccat atctggtggt ggatgccaca tacctgggcg
     1261 tgaacgtgcc cgtggagtat gtgaaagacg gtcagatcgt gctgaatctg tctgcaagtg
     1321 cgaccggcaa cctgcaactg acaaatgatt ttatccagtt caacgcccgc tttaagggcg
     1381 tgtctcgtga actgtatatc ccgatgggtg ccgctctggc catttacgct cgcgagaacg
     1441 gcgatggtgt gatgttcgaa ccagaagaaa tctatgacga gctgaatatt ggtccaccgg
     1501 tcgccaccat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
     1561 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     1621 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
     1681 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg
     1741 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     1801 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     1861 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     1921 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     1981 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
     2041 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
     2101 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     2161 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggactc agatctcgag
     2221 ctcaagcttc gaacgaattc ggcattgatc tgagcggcct gaccctgcag gaattcagct
     2281 ccccgaaacg ccctaagctg ctgcgtgaat attacgattg gctggttgat aacagcttta
     2341 ccccatatct ggtggtggat gccacatacc tgggcgtgaa cgtgcccgtg gagtatgtga
     2401 aagacggtca gatcgtgctg aatctgtctg caagtgcgac cggcaacctg caactgacaa
     2461 atgattttat ccagttcaac gcccgcttta agggcgtgtc tcgtgaactg tatatcccga
     2521 tgggtgccgc tctggccatt tacgctcgcg agaacggcga tggtgtgatg ttcgaaccag
     2581 aagaaatcta tgacgagctg aatattggtg gatccatggg actcagatct cgagctcaag
     2641 cttcgaacga attcggcatt gatctgagcg gcctgaccct gcaggtccgg aatatgcgac
     2701 agctgtcgga tgcggataag ctgcgcaagg tgatctgtga actgctggag actgaacgca
     2761 cctatgtgaa agacttaaac tgcctcatgg agagatacct gaagcccctt cagaaggaga
     2821 cctttctcac ccaggatgag cttgatgtac tttttggaaa tttaaccgaa atggtggagt
     2881 ttcaagtcga gttccttaag actctagaag atggagtaag actggtccct gacttggaaa
     2941 agctggagaa ggttgaccag ttcaagaaag tgctcttctc tctgggggga tccttcctgt
     3001 actacgcgga ccgcttcaag ctctacagtg ccttctgtgc gagccacaca aaagtcccca
     3061 aggtcctggt gaaagccaag acggacacag ccttcaaggc gttcctcgat gcccagaacc
     3121 cgaggcagca gcactcatcc acgctggagt cttacctcat caagcccatc cagagggtcc
     3181 tcaagtaccc gcttctgctc agggagctgt ttgcgctgac cgatgcggag agcgaggaac
     3241 actaccacct ggacgtggcc atcaagacta tgaacaaggt cgccagtcac atcaatgaga
     3301 tgcagaagat ccacgaagag tttggtgctg tgtttgacca gctgattgct gagcagacgg
     3361 gagagaagaa agaggttgca gatctgagca tgggtgacct gctcttgcac accagcgtca
     3421 tctggctgaa cccacccgcc tcactgggaa agtggaaaaa ggagccagaa ttagcagcct
     3481 tcgtcttcaa aacggccgtg gtccttgtat ataaagacgg ttccaagcag aagaagaaac
     3541 ttgttggctc tcacaggctg tcaatctacg aggagtggga ccctttccgg tttcgccaca
     3601 tgatccctac cgaagctttg caggtccgag ctctgcccag tgcagatgca gaggcaaatg
     3661 cggtatgcga aattgtccac gtgaaatcag agtcagaagg gaggcccgag cgggttttcc
     3721 acctctgctg cagttcccca gagagcagaa aggactttct gaagtctgtg cattcgatcc
     3781 tgcgagataa acacagaaga cagctcctca aaacggaaga attctgacaa ttgttgttgt
     3841 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3901 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3961 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4021 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4081 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4141 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4201 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4261 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4321 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4381 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     4441 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4501 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     4561 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4621 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4681 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4741 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4801 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4861 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     4921 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4981 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5041 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5101 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     5161 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5221 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5281 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5341 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5401 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5461 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5521 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5581 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5641 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5701 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5761 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5821 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5881 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     5941 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     6001 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     6061 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     6121 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     6181 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     6241 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6301 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6361 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6421 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6481 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6541 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6601 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6661 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6721 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6781 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6841 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6901 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6961 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7021 tgattctgtg gataaccgta ttaccgccat gcat
//
