LOCUS       sequence-153121-        6476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For gateway cloning Magneto2.0 into destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6476)
  AUTHORS   Wheeler MA, Smith CJ, Ottolini M, Barker BS, Purohit AM, Grippo RM, 
            Gaykema RP, Spano AJ, Beenhakker MP, Kucenas S, Patel MK, Deppmann 
            CD, Guler AD
  TITLE     Genetically targeted magnetic control of the nervous system.
  JOURNAL   Nat Neurosci. 2016 Mar 7. doi: 10.1038/nn.4265.
  PUBMED    26950006
REFERENCE   2  (bases 1 to 6476)
  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: "Nat 
            Neurosci. 2016 Mar 7. doi: 10.1038/nn.4265."
COMMENT     Sequence Label: pCR8-Magneto2.0
FEATURES             Location/Qualifiers
     source          1..6476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      692..701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(698..2974,2975..2995,2996..3019,3020..3049,3050..3568,
                     3569..3589,3590..3613,3614..3637,3638..4186,4187..4207,
                     4208..4231,4232..4261,4262..4321)
                     /codon_start=1
                     /product="fusion between a C-terminally truncated rat TRPV4
                     cation channel, the human ferritin light chain, the human 
                     ferritin heavy chain, and a plasma membrane targeting 
                     signal from the Kir2.1 potassium channel (Wheeler et al., 
                     2016)"
                     /label=Magneto2.0
                     /note="can be used as a magnetically sensitive actuator of 
                     neuronal activity"
                     /translation="MADPGDGPRAAPGDVAEPPGDESGTSGGEAFPLSSLANLFEGEEG
                     SSSLSPVDASRPAGPGDGRPNLRMKFQGAFRKGVPNPIDLLESTLYESSVVPGPKKAPM
                     DSLFDYGTYRHHPSDNKRWRRKVVEKQPQSPKAPAPQPPPILKVFNRPILFDIVSRGST
                     ADLDGLLSYLLTHKKRLTDEEFREPSTGKTCLPKALLNLSNGRNDTIPVLLDIAERTGN
                     MREFINSPFRDIYYRGQTALHIAIERRCKHYVELLVAQGADVHAQARGRFFQPKDEGGY
                     FYFGELPLSLAACTNQPHIVNYLTENPHKKADMRRQDSRGNTVLHALVAIADNTRENTK
                     FVTKMYDLLLLKCSRLFPDSNLETVLNNDGLSPLMMAAKTGKIGVFQHIIRREVTDEDT
                     RHLSRKFKDWAYGPVYSSLYDLSSLDTCGEEVSVLEILVYNSKIENRHEMLAVEPINEL
                     LRDKWRKFGAVSFYINVVSYLCAMVIFTLTAYYQPLEGTPPYPYRTTVDYLRLAGEVIT
                     LLTGVLFFFTSIKDLFMKKCPGVNSLFVDGSFQLLYFIYSVLVVVSAALYLAGIEAYLA
                     VMVFALVLGWMNALYFTRGLKLTGTYSIMIQKILFKDLFRFLLVYLLFMIGYASALVTL
                     LNPCTNMKVCNEDQSNCTVPSYPACRDSETFSAFLLDLFKLTIGMGDLEMLSSAKYPVV
                     FILLLVTYIILTFVLLLNMLIALMGETVGQVSKESKHIWKLQWATTILDIERSFPVFLR
                     KAFRSGARGGGGSDYKDDDDKGGGGSSRVMGSQIRQNYSTDVEAAVNSLVNLYLQASYT
                     YLSLGFYFDRDDVALEGVSHFFRELAEEKREGYERLLKMQNQRGGRALFQDIKKPAEDE
                     WGKTPDAMKAAMALEKKLNQALLDLHALGSARTDPHLCDFLETHFLDEEVKLIKKMGDH
                     LTNLHRLGGPEAGLGEYLFERLTLKHDARGGGGSDYKDDDDKGGGGSSRVMTTASTSQV
                     RQNYHQDSEAAINRQINLELYASYVYLSMSYYFDRDDVALKNFAKYFLHQSHEEREHAE
                     KLMKLQNQRGGRIFLQDIKKPDCDDWESGLNAMECALHLEKNVNQSLLELHKLATDKND
                     PHLCDFIETHYLNEQVKAIKELGDHVTNLRKMGAPESGLAEYLFDKHTLGDSDNESARG
                     GGGSDYKDDDDKGGGGSSRVASKSRITSEGEYIPLDQIDINV"
     protein_bind    complement(4355..4454)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4471..4490)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4472..4490)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4495..4511)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4495..4511)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4942..5733
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      5826..6414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6315..6334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gaattcgccc ttaaaggtac cgccaccatg gcagatcctg gtgatggccc
      721 ccgtgcagcg cctggggatg tggctgagcc ccctggagac gagagtggca cttctggtgg
      781 ggaggccttc cccctctctt ccctggccaa cctgtttgag ggagaggaag gctcctcttc
      841 tctttcacca gtggatgcta gccgccctgc tggccccggg gatggacgtc caaacctgcg
      901 tatgaagttc cagggcgctt tccgcaaggg ggttcccaac cccattgacc tgctggagtc
      961 caccctgtat gagtcctcag tagtgcctgg gcccaagaaa gcgcccatgg attcgttgtt
     1021 cgactatggc acttaccggc accaccccag tgacaacaag agatggagga ggaaggtcgt
     1081 agagaagcag ccacagagcc ccaaagctcc cgccccccag ccacccccca tcctcaaagt
     1141 cttcaaccgg cccatcctct ttgacatcgt gtcccggggc tccactgccg acctggacgg
     1201 actgctctcc tacttgctga cccacaagaa gcgcctgact gatgaggagt tccgggaacc
     1261 atccacaggg aagacctgcc tgcccaaggc acttctgaac ttaagcaatg gccgaaacga
     1321 caccatccca gtgttgctgg acattgcgga acgcacgggc aacatgcggg agttcatcaa
     1381 ctcgcccttc agagacatct actaccgagg gcagacggca ctgcacatcg ccattgaacg
     1441 gcgctgcaag cattacgtgg agctcctggt ggcccaggga gccgatgtgc acgcgcaggc
     1501 ccgagggcgg ttcttccagc ccaaggatga gggtggctac ttctactttg gggagctgcc
     1561 cttgtccttg gcagcctgca ccaaccagcc gcacatcgtc aactacctga cagagaaccc
     1621 tcacaagaaa gccgatatga ggcgacagga ctccagaggc aacacggtgc tccacgcgct
     1681 ggtggccatc gctgacaaca cccgagagaa caccaagttt gtcaccaaga tgtatgacct
     1741 gttgcttctc aagtgctccc gcctcttccc agacagcaac ctggagactg tgcttaacaa
     1801 tgacggtctt tcgcccctca tgatggctgc caagactggc aagatcgggg tctttcagca
     1861 catcatccga cgggaggtga cagatgagga cacacggcac ctgtctcgca agttcaagga
     1921 ctgggcctac gggcctgtgt attcttctct ctacgacctc tcctccctgg atacgtgcgg
     1981 ggaggaagtg tccgtgctgg agatcctggt ttacaacagc aagatcgaga accgccatga
     2041 gatgctggct gtggagccca ttaacgaact gctgagggac aagtggcgta agttcggggc
     2101 cgtgtccttc tacatcaacg ttgtctccta tctgtgtgcc atggtcatct tcaccctcac
     2161 agcctactat cagccactgg agggcacgcc accctaccct taccgtacca cggtggacta
     2221 cctgaggctg gctggtgagg tcatcacgct cctcacagga gtcctgttct tctttaccag
     2281 tatcaaagac ttgttcatga agaaatgccc tggagtgaat tctctcttcg tcgatggctc
     2341 cttccagttg ctctacttca tctactcagt gctggtggtt gtgtctgcgg cgctctacct
     2401 ggcagggatc gaggcctatc tggctgtgat ggtctttgcc ctggtcctgg gctggatgaa
     2461 tgccctttac ttcacccgtg ggctgaagct gacagggacc tacagcatca tgattcagaa
     2521 gatcctcttc aaagatctct tccgctttct gctggtctac ctgcttttta tgattggcta
     2581 tgcctcagct ctggtcaccc tcctgaatcc gtgcaccaac atgaaggtct gtaacgagga
     2641 ccagagcaac tgcacggtgc cctcataccc cgcgtgccgg gacagcgaga ccttcagcgc
     2701 cttcctactg gacctcttca agctcaccat cggcatgggc gacctggaga tgctgagcag
     2761 cgctaagtac cccgtggtct tcattctcct gctggttacc tacatcatcc tcaccttcgt
     2821 gctcctgctg aacatgctca tcgccctcat gggtgagacc gtgggccagg tgtccaagga
     2881 gagcaagcac atctggaagc tgcagtgggc caccaccatc ctggacatcg agcgctcctt
     2941 ccctgtgttc ctgaggaagg ccttccgctc cggagcacgg ggcgggggcg gttctgatta
     3001 taaggacgac gacgacaaag gaggaggcgg ctcctctcgg gttatgggct cccagattcg
     3061 tcagaattat tccaccgacg tggaggcagc cgtcaacagc ctggtcaatt tgtacctgca
     3121 ggcctcctac acctacctct ctctgggctt ctatttcgac cgcgatgatg tggctctgga
     3181 aggcgtgagc cacttcttcc gcgaattggc cgaggagaag cgcgagggct acgagcgtct
     3241 cctgaagatg caaaaccagc gtggcggccg cgctctcttc caggacatca agaagccagc
     3301 tgaagatgag tggggtaaaa ccccagacgc catgaaagct gccatggccc tggagaaaaa
     3361 gctgaaccag gcccttttgg atcttcatgc cctgggttct gcccgcacgg acccccatct
     3421 ctgtgacttc ctggagactc acttcctaga tgaggaagtg aagcttatca agaagatggg
     3481 tgaccacctg accaacctcc acaggctggg tggcccggag gctgggctgg gcgagtatct
     3541 cttcgaaagg ctcactctca agcacgacgc ccgcggcggc ggcggctccg attataaaga
     3601 tgatgatgat aaaggcggcg gcggctcctc ccgcgtcatg acgaccgcgt ccacctcgca
     3661 ggtgcgccag aactaccacc aggactcaga ggccgccatc aaccgccaga tcaacctgga
     3721 gctctacgcc tcctacgttt acctgtccat gtcttactac tttgaccgcg atgatgtggc
     3781 tttgaagaac tttgccaaat actttcttca ccaatctcat gaggagaggg aacatgctga
     3841 gaaactgatg aagctgcaga accaacgagg tggccgaatc ttccttcagg atatcaagaa
     3901 accagactgt gatgactggg agagcgggct gaatgcaatg gagtgtgcat tacatttgga
     3961 aaaaaatgtg aatcagtcac tactggaact gcacaaactg gccactgaca aaaatgaccc
     4021 ccatttgtgt gacttcattg agacacatta cctgaatgag caggtgaaag ccatcaaaga
     4081 attgggtgac cacgtgacca acttgcgcaa gatgggagcg cccgaatctg gcttggcgga
     4141 atatctcttt gacaagcaca ccctgggaga cagtgataat gaaagcgcac gcgggggtgg
     4201 cgggagcgac tataaagacg acgatgacaa agggggtgga ggctcctccc gagtggctag
     4261 caagagcaga atcaccagcg agggcgagta catccccctg gaccagatcg acatcaacgt
     4321 gtaaaccggt ggatccgcta gaagggcgaa ttcgacccag ctttcttgta caaagttggc
     4381 attataaaaa ataattgctc atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     4441 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     4501 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     4561 aaaaatatat catcatgcct cctctagacc agccaggaca gaaatgcctc gacttcgctg
     4621 ctgcccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg
     4681 acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg
     4741 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
     4801 tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac
     4861 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc
     4921 cctaaaacaa agttaaacat catgagggaa gcggtgatcg ccgaagtatc gactcaacta
     4981 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
     5041 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg
     5101 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
     5161 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
     5221 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
     5281 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
     5341 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
     5401 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
     5461 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
     5521 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
     5581 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
     5641 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
     5701 aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc
     5761 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5821 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5881 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5941 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     6001 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6061 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6121 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6181 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     6241 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6301 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6361 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6421 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
