LOCUS       sequence-224716-        6572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  anchor iLID domain to plasma membrane with slower diffusion .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6572)
  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 6572)
  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: iLID-eGFP-2xC2-iLID
FEATURES             Location/Qualifiers
     source          1..6572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             245..682
                     /codon_start=1
                     /product="improved light-inducible dimer when used with 
                     SspB nano or SspB micro (Guntas et al., 2015)"
                     /label=iLID
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGDVQ
                     YFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     regulatory      692..701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(698..700,701..703,704..1414)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(698..700,701..703,704..1414)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(743..764)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1004..1023)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1351..1372
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2414..2851
                     /codon_start=1
                     /product="improved light-inducible dimer when used with 
                     SspB nano or SspB micro (Guntas et al., 2015)"
                     /label=iLID
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGDVQ
                     YFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     polyA_signal    2996..3117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3033..3052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3087..3106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3124..3579)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3261..3282)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3473..3492
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3606..3710
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3712..4069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3712..3903
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3735..3755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3920..4055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4104..4898
                     /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             4104..4898
                     /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(4158..4177)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4768..4787
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5086..5105)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5130..5177
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5506..6094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5995..6014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6269..6572
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcatggggga
      241 gtttctggca accacactgg aacggatcga gaaaaatttc gtgattactg atccgagact
      301 gcctgacaac ccaatcattt ttgcgagcga ttccttcctg cagctgacag aatattctcg
      361 ggaagagatc ctggggcgca attgccgttt tctgcaggga cccgagacag accgtgccac
      421 tgttcggaaa atcagagatg ctattgacaa ccagactgaa gtgaccgttc agctgatcaa
      481 ttataccaag agcggcaaga agttctggaa cgtgttccac ctgcagccga tgcgcgatta
      541 taagggcgac gtccagtact tcattggcgt gcagctggat ggcaccgaac gtcttcatgg
      601 cgccgctgag cgtgaggcgg tctgcctgat caaaaagaca gcctttcaga ttgctgaggc
      661 agcgaacgac gaaaattact ttggaccggt cgccaccatg gtgagcaagg gcgaggagct
      721 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      781 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
      841 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
      901 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      961 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1021 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1081 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1141 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1201 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1261 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1321 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1381 cgggatcact ctcggcatgg acgagctgta caagtccgga agtagttgca ctgaacccct
     1441 aggcctgaag gataatacca tccccaacaa gcagatcaca gcctccagct actacaaaac
     1501 ctggggcctg agtgccttta gctggtttcc ctactacgca cgactggata atcagggcaa
     1561 gttcaacgcc tggaccgccc agaccaacag tgcctctgag tggctgcaga ttgacctggg
     1621 ctcccagaag cgagtcacgg gcatcatcac ccagggtgcc cgagactttg gccacattca
     1681 atatgtggct gcctacaggg tggcctatgg tgatgatggt gtgacctgga ctgagtacaa
     1741 ggacccgggg gcctcagaaa gcaagatttt ccctggtaac atggacaata attcccacaa
     1801 gaagaacata tttgagacgc cgttccaggc tcgcttcgtg cggatccagc ccgtggcctg
     1861 gcacaaccgt atcaccctgc gagtggagct gctgggctgt ggtaagctta gatcttgcac
     1921 tgaaccccta ggcctgaagg ataataccat ccccaacaag cagatcacag cctccagcta
     1981 ctacaaaacc tggggcctga gtgcctttag ctggtttccc tactacgcac gactggataa
     2041 tcagggcaag ttcaacgcct ggaccgccca gaccaacagt gcctctgagt ggctgcagat
     2101 tgacctgggc tcccagaagc gagtcacggg catcatcacc cagggtgccc gagactttgg
     2161 ccacattcaa tatgtggctg cctacagggt ggcctatggt gatgatggtg tgacctggac
     2221 tgagtacaag gacccggggg cctcagaaag caagattttc cctggtaaca tggacaataa
     2281 ttcccacaag aagaacatat ttgagacgcc gttccaggct cgcttcgtgc ggatccagcc
     2341 cgtggcctgg cacaaccgta tcaccctgcg agtggagctg ctgggctgtt atgaattcag
     2401 catgggggag tttctggcaa ccacactgga acggatcgag aaaaatttcg tgattactga
     2461 tccgagactg cctgacaacc caatcatttt tgcgagcgat tccttcctgc agctgacaga
     2521 atattctcgg gaagagatcc tggggcgcaa ttgccgtttt ctgcagggac ccgagacaga
     2581 ccgtgccact gttcggaaaa tcagagatgc tattgacaac cagactgaag tgaccgttca
     2641 gctgatcaat tataccaaga gcggcaagaa gttctggaac gtgttccacc tgcagccgat
     2701 gcgcgattat aagggcgacg tccagtactt cattggcgtg cagctggatg gcaccgaacg
     2761 tcttcatggc gccgctgagc gtgaggcggt ctgcctgatc aaaaagacag cctttcagat
     2821 tgctgaggca gcgaacgacg aaaattactt tggtaccgcg ggcccgggat ccaccggatc
     2881 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2941 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3001 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3061 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     3121 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3181 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3241 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3301 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3361 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3421 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3481 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3541 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3601 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3661 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3721 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3781 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3841 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3901 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3961 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4021 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4081 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4141 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4201 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4261 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4321 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4381 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4441 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4501 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4561 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4621 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4681 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4741 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4801 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4861 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4921 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4981 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5041 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5101 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5161 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5221 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5281 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5341 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5401 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5461 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5521 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5581 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5641 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5701 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5761 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5821 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5881 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5941 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6001 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6061 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6121 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6181 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
     6241 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     6301 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     6361 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     6421 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     6481 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     6541 acatctacgt attagtcatc gctattacca tg
//
