LOCUS       sequence-474956-        7448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of Magneto2.0 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7448)
  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 7448)
  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: pAAV-CMV-DIO-Magneto2.0-sNRPpA
FEATURES             Location/Qualifiers
     source          1..7448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        196..499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     654..674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    complement(766..799)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(850..883)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(898..957,958..987,988..1011,1012..1032,
                     1033..1581,1582..1605,1606..1629,1630..1650,1651..2169,
                     2170..2199,2200..2223,2224..2244,2245..4521))
                     /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"
     regulatory      4518..4527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4540..4573
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    4624..4657
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   4711..4851
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4711..4840
                     /label=AAV2 ITR
     rep_origin      4926..5381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5013..5032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5223..5244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5398..5417)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5517..5539
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5577..5595)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5663..5767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5768..5836,5837..6628)
                     /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     complement(5986..6005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6799..7387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7288..7307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgcctagta atcaattacg gggtcattag ttcatagccc
      181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      361 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      421 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      481 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      541 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      601 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      661 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      721 acccactgct tactggctta tcgaaattct agagtcgact ccggaataac ttcgtatagg
      781 atactttata cgaagttatg cagaatggta gctggattgt agctgctatt agcaatatga
      841 aacctcttaa taacttcgta tagcatacat tatacgaagt tatggcgcgc cggatcccac
      901 gttgatgtcg atctggtcca gggggatgta ctcgccctcg ctggtgattc tgctcttgct
      961 agccactcgg gaggagcctc cacccccttt gtcatcgtcg tctttatagt cgctcccgcc
     1021 acccccgcgt gcgctttcat tatcactgtc tcccagggtg tgcttgtcaa agagatattc
     1081 cgccaagcca gattcgggcg ctcccatctt gcgcaagttg gtcacgtggt cacccaattc
     1141 tttgatggct ttcacctgct cattcaggta atgtgtctca atgaagtcac acaaatgggg
     1201 gtcatttttg tcagtggcca gtttgtgcag ttccagtagt gactgattca catttttttc
     1261 caaatgtaat gcacactcca ttgcattcag cccgctctcc cagtcatcac agtctggttt
     1321 cttgatatcc tgaaggaaga ttcggccacc tcgttggttc tgcagcttca tcagtttctc
     1381 agcatgttcc ctctcctcat gagattggtg aagaaagtat ttggcaaagt tcttcaaagc
     1441 cacatcatcg cggtcaaagt agtaagacat ggacaggtaa acgtaggagg cgtagagctc
     1501 caggttgatc tggcggttga tggcggcctc tgagtcctgg tggtagttct ggcgcacctg
     1561 cgaggtggac gcggtcgtca tgacgcggga ggagccgccg ccgcctttat catcatcatc
     1621 tttataatcg gagccgccgc cgccgcgggc gtcgtgcttg agagtgagcc tttcgaagag
     1681 atactcgccc agcccagcct ccgggccacc cagcctgtgg aggttggtca ggtggtcacc
     1741 catcttcttg ataagcttca cttcctcatc taggaagtga gtctccagga agtcacagag
     1801 atgggggtcc gtgcgggcag aacccagggc atgaagatcc aaaagggcct ggttcagctt
     1861 tttctccagg gccatggcag ctttcatggc gtctggggtt ttaccccact catcttcagc
     1921 tggcttcttg atgtcctgga agagagcgcg gccgccacgc tggttttgca tcttcaggag
     1981 acgctcgtag ccctcgcgct tctcctcggc caattcgcgg aagaagtggc tcacgccttc
     2041 cagagccaca tcatcgcggt cgaaatagaa gcccagagag aggtaggtgt aggaggcctg
     2101 caggtacaaa ttgaccaggc tgttgacggc tgcctccacg tcggtggaat aattctgacg
     2161 aatctgggag cccataaccc gagaggagcc gcctcctcct ttgtcgtcgt cgtccttata
     2221 atcagaaccg cccccgcccc gtgctccgga gcggaaggcc ttcctcagga acacagggaa
     2281 ggagcgctcg atgtccagga tggtggtggc ccactgcagc ttccagatgt gcttgctctc
     2341 cttggacacc tggcccacgg tctcacccat gagggcgatg agcatgttca gcaggagcac
     2401 gaaggtgagg atgatgtagg taaccagcag gagaatgaag accacggggt acttagcgct
     2461 gctcagcatc tccaggtcgc ccatgccgat ggtgagcttg aagaggtcca gtaggaaggc
     2521 gctgaaggtc tcgctgtccc ggcacgcggg gtatgagggc accgtgcagt tgctctggtc
     2581 ctcgttacag accttcatgt tggtgcacgg attcaggagg gtgaccagag ctgaggcata
     2641 gccaatcata aaaagcaggt agaccagcag aaagcggaag agatctttga agaggatctt
     2701 ctgaatcatg atgctgtagg tccctgtcag cttcagccca cgggtgaagt aaagggcatt
     2761 catccagccc aggaccaggg caaagaccat cacagccaga taggcctcga tccctgccag
     2821 gtagagcgcc gcagacacaa ccaccagcac tgagtagatg aagtagagca actggaagga
     2881 gccatcgacg aagagagaat tcactccagg gcatttcttc atgaacaagt ctttgatact
     2941 ggtaaagaag aacaggactc ctgtgaggag cgtgatgacc tcaccagcca gcctcaggta
     3001 gtccaccgtg gtacggtaag ggtagggtgg cgtgccctcc agtggctgat agtaggctgt
     3061 gagggtgaag atgaccatgg cacacagata ggagacaacg ttgatgtaga aggacacggc
     3121 cccgaactta cgccacttgt ccctcagcag ttcgttaatg ggctccacag ccagcatctc
     3181 atggcggttc tcgatcttgc tgttgtaaac caggatctcc agcacggaca cttcctcccc
     3241 gcacgtatcc agggaggaga ggtcgtagag agaagaatac acaggcccgt aggcccagtc
     3301 cttgaacttg cgagacaggt gccgtgtgtc ctcatctgtc acctcccgtc ggatgatgtg
     3361 ctgaaagacc ccgatcttgc cagtcttggc agccatcatg aggggcgaaa gaccgtcatt
     3421 gttaagcaca gtctccaggt tgctgtctgg gaagaggcgg gagcacttga gaagcaacag
     3481 gtcatacatc ttggtgacaa acttggtgtt ctctcgggtg ttgtcagcga tggccaccag
     3541 cgcgtggagc accgtgttgc ctctggagtc ctgtcgcctc atatcggctt tcttgtgagg
     3601 gttctctgtc aggtagttga cgatgtgcgg ctggttggtg caggctgcca aggacaaggg
     3661 cagctcccca aagtagaagt agccaccctc atccttgggc tggaagaacc gccctcgggc
     3721 ctgcgcgtgc acatcggctc cctgggccac caggagctcc acgtaatgct tgcagcgccg
     3781 ttcaatggcg atgtgcagtg ccgtctgccc tcggtagtag atgtctctga agggcgagtt
     3841 gatgaactcc cgcatgttgc ccgtgcgttc cgcaatgtcc agcaacactg ggatggtgtc
     3901 gtttcggcca ttgcttaagt tcagaagtgc cttgggcagg caggtcttcc ctgtggatgg
     3961 ttcccggaac tcctcatcag tcaggcgctt cttgtgggtc agcaagtagg agagcagtcc
     4021 gtccaggtcg gcagtggagc cccgggacac gatgtcaaag aggatgggcc ggttgaagac
     4081 tttgaggatg gggggtggct ggggggcggg agctttgggg ctctgtggct gcttctctac
     4141 gaccttcctc ctccatctct tgttgtcact ggggtggtgc cggtaagtgc catagtcgaa
     4201 caacgaatcc atgggcgctt tcttgggccc aggcactact gaggactcat acagggtgga
     4261 ctccagcagg tcaatggggt tgggaacccc cttgcggaaa gcgccctgga acttcatacg
     4321 caggtttgga cgtccatccc cggggccagc agggcggcta gcatccactg gtgaaagaga
     4381 agaggagcct tcctctccct caaacaggtt ggccagggaa gagaggggga aggcctcccc
     4441 accagaagtg ccactctcgt ctccaggggg ctcagccaca tccccaggcg ctgcacgggg
     4501 gccatcacca ggatctgcca tggtggcggt acctctagca taacttcgta taaagtatcc
     4561 tatacgaagt tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa
     4621 agaataactt cgtataatgt atgctatacg aagttatgaa ttcaaataaa atacgaaatg
     4681 cacgtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     4741 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4801 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     4861 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     4921 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4981 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     5041 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5101 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5161 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5221 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     5281 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     5341 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5401 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5461 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5521 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5581 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5641 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5701 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5761 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5821 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5881 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5941 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6001 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6061 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6121 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6181 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6241 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6301 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6361 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6421 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6481 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6541 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6601 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6661 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6721 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6781 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6841 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6901 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6961 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7021 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7081 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7141 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7201 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7261 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7321 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7381 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7441 tcacatgt
//
