LOCUS       sequence-215992-        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"
     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"
     CDS             609..1046
                     /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      1056..1065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1062..1064,1065..1067,1068..1778)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1062..1064,1065..1067,1068..1778)
                     /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(1107..1128)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1368..1387)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1715..1736
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2778..3215
                     /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    3360..3481
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3397..3416)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3451..3470
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3488..3943)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3625..3646)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3837..3856
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3970..4074
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4076..4433
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4076..4267
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4099..4119)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4284..4419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4468..5262
                     /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             4468..5262
                     /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(4522..4541)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5132..5151
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5450..5469)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5494..5541
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5870..6458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6359..6378
                     /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 gctagcatgg
      601 gggagtttct ggcaaccaca ctggaacgga tcgagaaaaa tttcgtgatt actgatccga
      661 gactgcctga caacccaatc atttttgcga gcgattcctt cctgcagctg acagaatatt
      721 ctcgggaaga gatcctgggg cgcaattgcc gttttctgca gggacccgag acagaccgtg
      781 ccactgttcg gaaaatcaga gatgctattg acaaccagac tgaagtgacc gttcagctga
      841 tcaattatac caagagcggc aagaagttct ggaacgtgtt ccacctgcag ccgatgcgcg
      901 attataaggg cgacgtccag tacttcattg gcgtgcagct ggatggcacc gaacgtcttc
      961 atggcgccgc tgagcgtgag gcggtctgcc tgatcaaaaa gacagccttt cagattgctg
     1021 aggcagcgaa cgacgaaaat tactttggac cggtcgccac catggtgagc aagggcgagg
     1081 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1141 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1201 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1261 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1321 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1381 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1441 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1501 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1561 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1621 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1681 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1741 ccgccgggat cactctcggc atggacgagc tgtacaagtc cggaagtagt tgcactgaac
     1801 ccctaggcct gaaggataat accatcccca acaagcagat cacagcctcc agctactaca
     1861 aaacctgggg cctgagtgcc tttagctggt ttccctacta cgcacgactg gataatcagg
     1921 gcaagttcaa cgcctggacc gcccagacca acagtgcctc tgagtggctg cagattgacc
     1981 tgggctccca gaagcgagtc acgggcatca tcacccaggg tgcccgagac tttggccaca
     2041 ttcaatatgt ggctgcctac agggtggcct atggtgatga tggtgtgacc tggactgagt
     2101 acaaggaccc gggggcctca gaaagcaaga ttttccctgg taacatggac aataattccc
     2161 acaagaagaa catatttgag acgccgttcc aggctcgctt cgtgcggatc cagcccgtgg
     2221 cctggcacaa ccgtatcacc ctgcgagtgg agctgctggg ctgtggtaag cttagatctt
     2281 gcactgaacc cctaggcctg aaggataata ccatccccaa caagcagatc acagcctcca
     2341 gctactacaa aacctggggc ctgagtgcct ttagctggtt tccctactac gcacgactgg
     2401 ataatcaggg caagttcaac gcctggaccg cccagaccaa cagtgcctct gagtggctgc
     2461 agattgacct gggctcccag aagcgagtca cgggcatcat cacccagggt gcccgagact
     2521 ttggccacat tcaatatgtg gctgcctaca gggtggccta tggtgatgat ggtgtgacct
     2581 ggactgagta caaggacccg ggggcctcag aaagcaagat tttccctggt aacatggaca
     2641 ataattccca caagaagaac atatttgaga cgccgttcca ggctcgcttc gtgcggatcc
     2701 agcccgtggc ctggcacaac cgtatcaccc tgcgagtgga gctgctgggc tgttatgaat
     2761 tcagcatggg ggagtttctg gcaaccacac tggaacggat cgagaaaaat ttcgtgatta
     2821 ctgatccgag actgcctgac aacccaatca tttttgcgag cgattccttc ctgcagctga
     2881 cagaatattc tcgggaagag atcctggggc gcaattgccg ttttctgcag ggacccgaga
     2941 cagaccgtgc cactgttcgg aaaatcagag atgctattga caaccagact gaagtgaccg
     3001 ttcagctgat caattatacc aagagcggca agaagttctg gaacgtgttc cacctgcagc
     3061 cgatgcgcga ttataagggc gacgtccagt acttcattgg cgtgcagctg gatggcaccg
     3121 aacgtcttca tggcgccgct gagcgtgagg cggtctgcct gatcaaaaag acagcctttc
     3181 agattgctga ggcagcgaac gacgaaaatt actttggtac cgcgggcccg ggatccaccg
     3241 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3301 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3361 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3421 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3481 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3541 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3601 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3661 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3721 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3781 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3841 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3901 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     3961 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4021 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     4081 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4141 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4201 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4261 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4321 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4381 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     4441 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4501 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4561 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4621 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     4681 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4741 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4801 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4861 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4921 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4981 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5041 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5101 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5161 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5221 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5281 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5341 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5401 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     5461 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     5521 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     5581 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     5641 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     5701 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5761 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5821 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5881 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5941 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6001 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6061 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6121 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6181 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6241 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6301 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6361 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6421 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6481 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6541 attctgtgga taaccgtatt accgccatgc at
//
