LOCUS       sequence-224590-        5954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Translocating to iLID domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5954)
  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 5954)
  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
FEATURES             Location/Qualifiers
     source          1..5954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             76..405
                     /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      418..427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             424..1119
                     /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(557..575)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     825..844
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1192..1521
                     /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    1657..1778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1694..1713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1748..1767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1785..2240)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1922..1943)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2134..2153
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2267..2371
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2373..2730
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2373..2564
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2396..2416)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2581..2716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2765..3559
                     /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             2765..3559
                     /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(2819..2838)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3429..3448
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3747..3766)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3791..3838
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4167..4755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4656..4675
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4930..5233
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5234..5437
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5388..5408
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    5478..5534
                     /label=MCS
                     /note="multiple cloning site"
     CDS             5610..5939
                     /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"
ORIGIN
        1 atgggactca gatctcgagc tcaagcttcg aacgaattcg gcattgatct gagcggcctg
       61 accctgcagg aattcagctc cccgaaacgc cctaagctgc tgcgtgaata ttacgattgg
      121 ctggttgata acagctttac cccatatctg gtggtggatg ccacatacct gggcgtgaac
      181 gtgcccgtgg agtatgtgaa agacggtcag atcgtgctga atctgtctgc aagtgcgacc
      241 ggcaacctgc aactgacaaa tgattttatc cagttcaacg cccgctttaa gggcgtgtct
      301 cgtgaactgt atatcccgat gggtgccgct ctggccattt acgctcgcga gaacggcgat
      361 ggtgtgatgt tcgaaccaga agaaatctat gacgagctga atattggtcc accggtcgcc
      421 accatggtga gcaagggcga ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg
      481 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      541 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttctcctgg
      601 gacatcctgt cccctcagtt catgtacggc tccagggcct tcaccaagca ccccgccgac
      661 atccccgact actataagca gtccttcccc gagggcttca agtgggagcg cgtgatgaac
      721 ttcgaggacg gcggcgccgt gaccgtgacc caggacacct ccctggagga cggcaccctg
      781 atctacaagg tgaagctccg cggcaccaac ttccctcctg acggccccgt aatgcagaag
      841 aagacaatgg gctgggaagc gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag
      901 ggcgacatta agatggccct gcgcctgaag gacggcggcc gctacctggc ggacttcaag
      961 accacctaca aggccaagaa gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag
     1021 ttggacatca cctcccacaa cgaggactac accgtggtgg aacagtacga acgctccgag
     1081 ggccgccact ccaccggcgg catggacgag ctgtacaagg gactcagatc tcgagctcaa
     1141 gcttcgaacg aattcggcat tgatctgagc ggcctgaccc tgcaggaatt cagctccccg
     1201 aaacgcccta agctgctgcg tgaatattac gattggctgg ttgataacag ctttacccca
     1261 tatctggtgg tggatgccac atacctgggc gtgaacgtgc ccgtggagta tgtgaaagac
     1321 ggtcagatcg tgctgaatct gtctgcaagt gcgaccggca acctgcaact gacaaatgat
     1381 tttatccagt tcaacgcccg ctttaagggc gtgtctcgtg aactgtatat cccgatgggt
     1441 gccgctctgg ccatttacgc tcgcgagaac ggcgatggtg tgatgttcga accagaagaa
     1501 atctatgacg agctgaatat tggttaaaca tgtttgcggc cgcgactcta gatcataatc
     1561 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1621 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1681 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1741 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     1801 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     1861 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     1921 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     1981 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2041 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2101 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2161 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2221 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2281 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2341 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     2401 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2461 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2521 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2581 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2641 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2701 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     2761 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     2821 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     2881 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     2941 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3001 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3061 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3121 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3181 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3241 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     3301 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3361 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3421 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3481 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3541 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3601 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3661 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3721 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     3781 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     3841 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     3901 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     3961 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4021 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4081 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4141 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4201 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4261 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4321 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4381 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4441 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4501 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4561 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4621 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4681 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4741 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4801 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4861 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     4921 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     4981 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     5041 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     5101 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     5161 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     5221 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     5281 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     5341 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     5401 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
     5461 ctagcgctac cggactcaga tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg
     5521 cgggcccggg atccatggga ctcagatctc gagctcaagc ttcgaacgaa ttcggcattg
     5581 atctgagcgg cctgaccctg caggaattca gctccccgaa acgccctaag ctgctgcgtg
     5641 aatattacga ttggctggtt gataacagct ttaccccata tctggtggtg gatgccacat
     5701 acctgggcgt gaacgtgccc gtggagtatg tgaaagacgg tcagatcgtg ctgaatctgt
     5761 ctgcaagtgc gaccggcaac ctgcaactga caaatgattt tatccagttc aacgcccgct
     5821 ttaagggcgt gtctcgtgaa ctgtatatcc cgatgggtgc cgctctggcc atttacgctc
     5881 gcgagaacgg cgatggtgtg atgttcgaac cagaagaaat ctatgacgag ctgaatattg
     5941 gtccaccggg atcc
//
