LOCUS       sequence-224707-        5124 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 5124)
  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 5124)
  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: mSc-sspB
FEATURES             Location/Qualifiers
     source          1..5124
                     /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"
     misc_feature    227..307
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      309..318
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             315..1010
                     /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(448..466)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     716..735
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1083..1412
                     /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    1548..1669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1585..1604)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1639..1658
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1676..2131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1813..1834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2025..2044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2158..2262
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2264..2621
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2264..2455
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2287..2307)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2472..2607
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2656..3450
                     /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             2656..3450
                     /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(2710..2729)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3320..3339
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3638..3657)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3682..3729
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4058..4646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4547..4566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4821..5124
                     /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 gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc
      301 caccggtcgc caccatggtg agcaagggcg aggcagtgat caaggagttc atgcggttca
      361 aggtgcacat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg
      421 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
      481 ccttctcctg ggacatcctg tcccctcagt tcatgtacgg ctccagggcc ttcaccaagc
      541 accccgccga catccccgac tactataagc agtccttccc cgagggcttc aagtgggagc
      601 gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc tccctggagg
      661 acggcaccct gatctacaag gtgaagctcc gcggcaccaa cttccctcct gacggccccg
      721 taatgcagaa gaagacaatg ggctgggaag cgtccaccga gcggttgtac cccgaggacg
      781 gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggc cgctacctgg
      841 cggacttcaa gaccacctac aaggccaaga agcccgtgca gatgcccggc gcctacaacg
      901 tcgaccgcaa gttggacatc acctcccaca acgaggacta caccgtggtg gaacagtacg
      961 aacgctccga gggccgccac tccaccggcg gcatggacga gctgtacaag ggactcagat
     1021 ctcgagctca agcttcgaac gaattcggca ttgatctgag cggcctgacc ctgcaggaat
     1081 tcagctcccc gaaacgccct aagctgctgc gtgaatatta cgattggctg gttgataaca
     1141 gctttacccc atatctggtg gtggatgcca catacctggg cgtgaacgtg cccgtggagt
     1201 atgtgaaaga cggtcagatc gtgctgaatc tgtctgcaag tgcgaccggc aacctgcaac
     1261 tgacaaatga ttttatccag ttcaacgccc gctttaaggg cgtgtctcgt gaactgtata
     1321 tcccgatggg tgccgctctg gccatttacg ctcgcgagaa cggcgatggt gtgatgttcg
     1381 aaccagaaga aatctatgac gagctgaata ttggttaaac atgtttgcgg ccgcgactct
     1441 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1501 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1561 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1621 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     1681 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1741 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     1801 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     1861 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     1921 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     1981 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2041 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2101 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2161 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2221 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2281 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2341 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2401 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2461 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2521 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2581 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     2641 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     2701 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     2761 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     2821 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     2881 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     2941 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3001 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3061 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3121 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3181 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3241 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3301 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3361 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3421 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3481 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3541 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3601 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     3661 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     3721 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     3781 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     3841 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     3901 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3961 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4021 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4081 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4141 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4201 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4261 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4321 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4381 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4441 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4501 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4561 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4621 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4681 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4741 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     4801 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     4861 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     4921 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     4981 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     5041 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     5101 gtattagtca tcgctattac catg
//
