LOCUS       sequence-140129-        7252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For gateway cloning Magneto2.0-p2A-mCherry into destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7252)
  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 7252)
  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-p2A-mCherry
FEATURES             Location/Qualifiers
     source          1..7252
                     /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"
     CDS             4337..4393
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4400..5107
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(4542..4560)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4810..4829
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(5131..5230)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5247..5266)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5248..5266)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5271..5287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5271..5287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5718..6509
                     /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      6602..7190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7091..7110
                     /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 gggatcaggc agcggcgcca cgaacttctc tctgttaaag caagcaggag acgtggaaga
     4381 aaaccccggt cccggatccg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     4441 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     4501 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     4561 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     4621 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     4681 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     4741 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     4801 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     4861 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     4921 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     4981 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     5041 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     5101 caagtaaggc gcgccaaaag ggcgaattcg acccagcttt cttgtacaaa gttggcatta
     5161 taaaaaataa ttgctcatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     5221 tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg
     5281 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     5341 atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctgc
     5401 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
     5461 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
     5521 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     5581 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     5641 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     5701 aaacaaagtt aaacatcatg agggaagcgg tgatcgccga agtatcgact caactatcag
     5761 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     5821 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     5881 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     5941 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     6001 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     6061 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     6121 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     6181 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     6241 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     6301 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     6361 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     6421 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     6481 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     6541 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6601 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6661 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6721 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6781 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6841 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6901 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6961 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     7021 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7081 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7141 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7201 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
