LOCUS       sequence-375649-        8268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses iLID-mCl3-MAP7N, a microtubule anchor used for optogenetic
            recruitment of SspB-mCh-p60, in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8268)
  AUTHORS   Meiring JCM, Grigoriev I, Nijenhuis W, Kapitein LC, Akhmanova A
  TITLE     Opto-katanin, an optogenetic tool for localized, microtubule 
            disassembly.
  JOURNAL   Curr Biol. 2022 Nov 7;32(21):4660-4674.e6. doi: 
            10.1016/j.cub.2022.09.010. Epub 2022 Sep 28.
  PUBMED    36174574
REFERENCE   2  (bases 1 to 8268)
  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"; doi: 
            "10.1016/j.cub.2022.09.010"; journalName: "Curr Biol"; date: 
            "2022-11-7- 7"; volume: "32"; issue: "21"; pages: "4660-4674.e6"
COMMENT     Sequence Label: iLID-mCl3-MAP7N
FEATURES             Location/Qualifiers
     source          1..8268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     941..963
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1035..1472
                     /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"
     CDS             join(1572..1574,1575..1577,1578..2288)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1617..1638)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2225..2246
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3426..3560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3463..3482)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3517..3536
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3574..3590)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3574..3590)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3587..3609)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3598..3614
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3622..3628,3629..3646,3647..3652))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3667..3688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3805..3822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3976..4564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4056..4075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4735..5526,5527..5595))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5358..5377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5596..5700)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5768..5786
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5824..5846)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5946..5965
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6159..6181
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6173..6190
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6174..6190
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7991..8266
                     /label=chicken beta-actin promoter
ORIGIN
        1 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
       61 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      121 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      181 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      241 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      301 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
      361 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
      421 gcgtgggggg gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac
      481 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tgcggggcgt
      541 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
      601 gggccgccyc gggccggggr gggctcgggg grggggcgcg gcggcccccg gagcgccggc
      661 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
      721 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
      781 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
      841 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccatc tccagcctcg gggctgtccg
      901 cagggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
      961 accggcgggg tttatatctt cccttctctg ttcctccgca gcccccaagc taggcaattg
     1021 ggcgcgccac catgctggca accacactgg aacggatcga gaaaaatttc gtgattactg
     1081 atccgagact gcctgacaac ccaatcattt ttgcgagcga ttccttcctg cagctgacag
     1141 aatattctcg ggaagagatc ctggggcgca attgccgttt tctgcaggga cccgagacag
     1201 accgtgccac tgttcggaaa atcagagatg ctattgacaa ccagactgaa gtgaccgttc
     1261 agctgatcaa ttataccaag agcggcaaga agttctggaa cgtgttccac ctgcagccga
     1321 tgcgcgatta taagggcgac gtccagtact tcattggcgt gcagctggat ggcaccgaac
     1381 gtcttcatgg cgccgctgag cgtgaggcgg tctgcctgat caaaaagaca gcctttcaga
     1441 ttgctgaggc agcgaacgac gaaaattact tttctagagg tggcgggggc agtgggggag
     1501 ggggaagtgg aggcggcggt tctggcgggg gaggctccgg tggcgggggg agcgggggtg
     1561 ggagtaagct tatggtgtct aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1621 tcgagctgga cggcgacgta aacggccaca agttcagcgt ccgcggcgag ggcgagggcg
     1681 atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1741 cctggcccac cctcgtgacc accttcggct acggcgtggc ctgcttcagc cgctaccccg
     1801 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1861 gcaccatctc tttcaaggac gacggtacct acaagacccg cgccgaggtg aagttcgagg
     1921 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1981 tcctggggca caagctggag tacaacttca acagccacta cgtctatatc acggccgaca
     2041 agcagaagaa ctgcatcaag gctaacttca agatccgcca caacgttgag gacggcagcg
     2101 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2161 ccgacaacca ctacctgagc catcagtcca agctgagcaa agaccccaac gagaagcgcg
     2221 atcacatggt cctgctggag ttcgtgaccg ccgccgggat tacacatggc atggacgagc
     2281 tgtacaaggc tagcggaggc ggtggctcag gcggaggagg gtcaggaggg ggaggcagtg
     2341 gcgggggcgg ctctggagga gggggttctg gtggtggtag ctacgtagcg gagctaggag
     2401 ctggcggcga cggccacagg ggcggcgacg gcgcagtgcg aagcgaaaca gcacccgaca
     2461 gctacaaagt gcaagataag aaaaatgcct ccagccgccc tgcctctgca atttcaggac
     2521 aaaataacaa ccactcagga aataaaccag accctccgcc tgtgttacgt gttgatgacc
     2581 ggcagcggct ggcccgggag cgacgtgagg aacgggagaa acagctagct gcaagagaaa
     2641 tagtgtggtt agaaagagaa gagcgagcca ggcagcacta cgagaagcac ctggaagagc
     2701 ggaagaagag gttggaggag cagaggcaga aggaggagcg gaggagggct gctgtggagg
     2761 agaagcggag gcagagactt gaggaggaca aagaacgcca cgaagctgtt gtacggcgca
     2821 caatggaaag gagccagaag ccaaaacaga agcataaccg ttggtcgtgg ggaggctctc
     2881 tccatgggag ccctagcatc cacagtgcag ctcgccgcct gcagctcagc ccatgggaga
     2941 gcagcgttgt taacagactc ctgacgccca cacattcgtt cctggccaga agtaaaagca
     3001 cagctgcctt gtctggagaa gcagcatctt gcagccccat catcatgccc tacaaagctg
     3061 cacactctag aaattcgatg gatcgaccaa aactctttgt aacaccacct gagggctctt
     3121 ctcgcaggag gatcattcat ggcacagcga gctataaaaa agaaagagag agagaaaatg
     3181 tactcttcct cacatctggc acccgaaggg ctgtatctcc atctaatccc aaagcaagac
     3241 aaccagctcg ctcccgactt tggcttccgt ccaagtctct tcctcatttg tgagtcgaca
     3301 ccggtcctag ataagtaatg atcataatca gccatatcac atctgtagag gttttacttg
     3361 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3421 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3481 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3541 tatcttatca tgtctggatc aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     3601 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3661 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3721 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3781 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3841 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3901 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3961 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4021 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4081 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4141 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4201 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4261 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4321 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4381 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4441 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4501 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4561 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4621 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4681 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4741 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4801 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4861 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4921 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4981 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5041 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5101 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5161 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5221 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5281 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5341 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5401 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5461 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5521 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5581 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5641 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5701 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5761 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5821 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5881 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     5941 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6001 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     6061 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6121 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     6181 gacggccagt gccaagttgg gatctttgca ttggcccacg gctctcagga tggggatgct
     6241 ccccttcagc acccggttcc ccttggaaac tgatggtcct ggctctgtgg catggcagtg
     6301 gcactgtgag gagcccccta ccagcagcac acagtggggt tggcactgcc acgctccgga
     6361 tgccgcgctc tgatccaacc ccataatcaa gggaacccga attgccccat cattgccccc
     6421 accaccccca tcctgccggg ccctcacacc ccacgctgcc ttctggtgac attccccagc
     6481 ccaaacccac ggcttcatgg ctaccgcggg gcatttccca ttgccgcccc attatcagct
     6541 ctgcacacct cccgctgtac ccatgcctcg tggctgccct tctttgacgt ataatcttct
     6601 aattaatacc cggccttgtc aaagtggagc acaaacgtta attaattccc cagcaggcag
     6661 gtaattaaca gtgtgactcc ctttttgctg cgagtggggc tgatacagag agatgtggca
     6721 ctatggagcc cacggggtcc tggcactggg tgcccacgga ggtccccatg tgctgcagtg
     6781 tcaccgcctc cgaggagaca gtattgtccc tgcggtgtcc ctgcagctca gctctgtcca
     6841 cagggccacc tccagtttgg aggggacaca atgcagcccc gatgcaaccc atcctcgcag
     6901 catcccaggg acaaagaccc cactgcaaga ccgcacacag ggctgggtcc cgctccccta
     6961 atatctacag tgcttcttgc atggcccctt aatcaatgca gttaatcagc atgcgctcat
     7021 gcaccgctct ggagctgcaa agcccctcgc agcgctgctc accaacaccg cgcaccgccc
     7081 cggcccagcc tgcagcacgc gctgcaaaca ggaaagaaac aaaatattgc ccaaatgtag
     7141 gcaaaggcat tcggctgcct tgacctccgc cgggccgggc cctgcctgac tcagctcctt
     7201 actcagcgct cgcttcctcc ctccggctgc caccgccgca gcgcacaccc tgacaaagag
     7261 tggcccttaa cgggctctga ggtgcaccca gcagtgcact cagcagtcca agggccggcc
     7321 tggaggtttg caccgctacg tgctgacatt agcattgaac ttggccctgg gtagtgctgc
     7381 aggccgggcg gggtgggtgt agagagtgca gcgcgcgttg cacccggtgc cccttcccct
     7441 cccttgcatc ccagcaggct gcaccccagc accaggcccg tgcatgcatg ctcctggtgt
     7501 tattgcagcc tggtgcatgc atgcgtcttg atggtgcagc gctgtgcatg catcctcctt
     7561 ggtgtgtagc agtctagtgc atgcataccc ctcggtgtta ttgctgctct gtgcacgcac
     7621 gctcattgta tcacttcatc ccagtgcatg cactcacact ggagcgattg ctgctcggtg
     7681 cacgcacact cattgtatca ctgcagctca gtggctgcac gcacaccggt gttattgctg
     7741 ctcggtgcgt gcatgcacat cagtgtcgct gcagctcagt gcatgcacgc tcattgccca
     7801 tcgctatccc tgcctctcct gctggcgctc cccgggaggt gacttcaagg ggaccgcagg
     7861 accacctcgg gggtgggggg agggctgcac acgcggaccc cgctccccct ccccaacaaa
     7921 gcactgtgga atcaaaaagg ggggaggggg gatggagggg cgcgtcacac ccccgcccca
     7981 caccctcacc tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     8041 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     8101 gggggggggg cgcgcgccag gsggggcggg gsggggsgrg gggsggggsg gggcgaggcg
     8161 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
     8221 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcggg
//
