LOCUS       sequence-224808-        6618 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 6618)
  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 6618)
  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-3xC2
FEATURES             Location/Qualifiers
     source          1..6618
                     /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"
     polyA_signal    3406..3527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3443..3462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3497..3516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3534..3989)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3671..3692)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3883..3902
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4016..4120
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4122..4479
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4122..4313
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4145..4165)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4330..4465
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4514..5308
                     /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             4514..5308
                     /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(4568..4587)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5178..5197
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5496..5515)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5540..5587
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5916..6504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6405..6424
                     /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 cttggatctt
     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 tgtggtaccg
     2761 gaagatcttg cactgaaccc ctaggcctga aggataatac catccccaac aagcagatca
     2821 cagcctccag ctactacaaa acctggggcc tgagtgcctt tagctggttt ccctactacg
     2881 cacgactgga taatcagggc aagttcaacg cctggaccgc ccagaccaac agtgcctctg
     2941 agtggctgca gattgacctg ggctcccaga agcgagtcac gggcatcatc acccagggtg
     3001 cccgagactt tggccacatt caatatgtgg ctgcctacag ggtggcctat ggtgatgatg
     3061 gtgtgacctg gactgagtac aaggacccgg gggcctcaga aagcaagatt ttccctggta
     3121 acatggacaa taattcccac aagaagaaca tatttgagac gccgttccag gctcgcttcg
     3181 tgcggatcca gcccgtggcc tggcacaacc gtatcaccct gcgagtggag ctgctgggct
     3241 gttaggaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg
     3301 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3361 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3421 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3481 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     3541 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3601 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3661 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3721 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3781 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3841 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3901 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3961 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4021 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4081 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4141 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4201 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4261 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4321 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4381 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4441 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4501 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4561 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4621 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4681 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4741 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4801 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4861 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4921 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4981 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5041 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5101 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5161 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5221 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5281 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5341 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5401 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5461 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5521 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5581 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5641 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5701 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5761 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5821 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5881 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5941 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6001 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6061 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6121 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6181 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6241 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6301 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6361 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6421 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6481 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6541 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6601 cgtattaccg ccatgcat
//
