LOCUS       Exported                7459 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Cre-dependent expression of Magneto2.0 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CMV-DIO-Magneto2.0-sNRPpA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7459)
  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 7459)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7459
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        207..510
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        511..714
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     665..685
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    777..810
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    861..894
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(909..4532)
                     /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      4529..4538
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(4551..4584)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(4635..4668)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     repeat_region   4722..4862
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4722..4851
                     /label=AAV2 ITR
     rep_origin      4937..5392
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5024..5043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5234..5255
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5409..5428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5528..5550
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5588..5606)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5674..5778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5779..6639
                     /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(5997..6016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6810..7398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7299..7318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgcctagt aatcaattac ggggtcatta
      181 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      241 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      301 ccaataggga ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg
      361 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      421 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      481 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      541 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      601 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      661 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
      721 ctaactagag aacccactgc ttactggctt atcgaaattc tagagtcgac tccggaataa
      781 cttcgtatag gatactttat acgaagttat gcagaatggt agctggattg tagctgctat
      841 tagcaatatg aaacctctta ataacttcgt atagcataca ttatacgaag ttatggcgcg
      901 ccggatccca cgttgatgtc gatctggtcc agggggatgt actcgccctc gctggtgatt
      961 ctgctcttgc tagccactcg ggaggagcct ccaccccctt tgtcatcgtc gtctttatag
     1021 tcgctcccgc cacccccgcg tgcgctttca ttatcactgt ctcccagggt gtgcttgtca
     1081 aagagatatt ccgccaagcc agattcgggc gctcccatct tgcgcaagtt ggtcacgtgg
     1141 tcacccaatt ctttgatggc tttcacctgc tcattcaggt aatgtgtctc aatgaagtca
     1201 cacaaatggg ggtcattttt gtcagtggcc agtttgtgca gttccagtag tgactgattc
     1261 acattttttt ccaaatgtaa tgcacactcc attgcattca gcccgctctc ccagtcatca
     1321 cagtctggtt tcttgatatc ctgaaggaag attcggccac ctcgttggtt ctgcagcttc
     1381 atcagtttct cagcatgttc cctctcctca tgagattggt gaagaaagta tttggcaaag
     1441 ttcttcaaag ccacatcatc gcggtcaaag tagtaagaca tggacaggta aacgtaggag
     1501 gcgtagagct ccaggttgat ctggcggttg atggcggcct ctgagtcctg gtggtagttc
     1561 tggcgcacct gcgaggtgga cgcggtcgtc atgacgcggg aggagccgcc gccgccttta
     1621 tcatcatcat ctttataatc ggagccgccg ccgccgcggg cgtcgtgctt gagagtgagc
     1681 ctttcgaaga gatactcgcc cagcccagcc tccgggccac ccagcctgtg gaggttggtc
     1741 aggtggtcac ccatcttctt gataagcttc acttcctcat ctaggaagtg agtctccagg
     1801 aagtcacaga gatgggggtc cgtgcgggca gaacccaggg catgaagatc caaaagggcc
     1861 tggttcagct ttttctccag ggccatggca gctttcatgg cgtctggggt tttaccccac
     1921 tcatcttcag ctggcttctt gatgtcctgg aagagagcgc ggccgccacg ctggttttgc
     1981 atcttcagga gacgctcgta gccctcgcgc ttctcctcgg ccaattcgcg gaagaagtgg
     2041 ctcacgcctt ccagagccac atcatcgcgg tcgaaataga agcccagaga gaggtaggtg
     2101 taggaggcct gcaggtacaa attgaccagg ctgttgacgg ctgcctccac gtcggtggaa
     2161 taattctgac gaatctggga gcccataacc cgagaggagc cgcctcctcc tttgtcgtcg
     2221 tcgtccttat aatcagaacc gcccccgccc cgtgctccgg agcggaaggc cttcctcagg
     2281 aacacaggga aggagcgctc gatgtccagg atggtggtgg cccactgcag cttccagatg
     2341 tgcttgctct ccttggacac ctggcccacg gtctcaccca tgagggcgat gagcatgttc
     2401 agcaggagca cgaaggtgag gatgatgtag gtaaccagca ggagaatgaa gaccacgggg
     2461 tacttagcgc tgctcagcat ctccaggtcg cccatgccga tggtgagctt gaagaggtcc
     2521 agtaggaagg cgctgaaggt ctcgctgtcc cggcacgcgg ggtatgaggg caccgtgcag
     2581 ttgctctggt cctcgttaca gaccttcatg ttggtgcacg gattcaggag ggtgaccaga
     2641 gctgaggcat agccaatcat aaaaagcagg tagaccagca gaaagcggaa gagatctttg
     2701 aagaggatct tctgaatcat gatgctgtag gtccctgtca gcttcagccc acgggtgaag
     2761 taaagggcat tcatccagcc caggaccagg gcaaagacca tcacagccag ataggcctcg
     2821 atccctgcca ggtagagcgc cgcagacaca accaccagca ctgagtagat gaagtagagc
     2881 aactggaagg agccatcgac gaagagagaa ttcactccag ggcatttctt catgaacaag
     2941 tctttgatac tggtaaagaa gaacaggact cctgtgagga gcgtgatgac ctcaccagcc
     3001 agcctcaggt agtccaccgt ggtacggtaa gggtagggtg gcgtgccctc cagtggctga
     3061 tagtaggctg tgagggtgaa gatgaccatg gcacacagat aggagacaac gttgatgtag
     3121 aaggacacgg ccccgaactt acgccacttg tccctcagca gttcgttaat gggctccaca
     3181 gccagcatct catggcggtt ctcgatcttg ctgttgtaaa ccaggatctc cagcacggac
     3241 acttcctccc cgcacgtatc cagggaggag aggtcgtaga gagaagaata cacaggcccg
     3301 taggcccagt ccttgaactt gcgagacagg tgccgtgtgt cctcatctgt cacctcccgt
     3361 cggatgatgt gctgaaagac cccgatcttg ccagtcttgg cagccatcat gaggggcgaa
     3421 agaccgtcat tgttaagcac agtctccagg ttgctgtctg ggaagaggcg ggagcacttg
     3481 agaagcaaca ggtcatacat cttggtgaca aacttggtgt tctctcgggt gttgtcagcg
     3541 atggccacca gcgcgtggag caccgtgttg cctctggagt cctgtcgcct catatcggct
     3601 ttcttgtgag ggttctctgt caggtagttg acgatgtgcg gctggttggt gcaggctgcc
     3661 aaggacaagg gcagctcccc aaagtagaag tagccaccct catccttggg ctggaagaac
     3721 cgccctcggg cctgcgcgtg cacatcggct ccctgggcca ccaggagctc cacgtaatgc
     3781 ttgcagcgcc gttcaatggc gatgtgcagt gccgtctgcc ctcggtagta gatgtctctg
     3841 aagggcgagt tgatgaactc ccgcatgttg cccgtgcgtt ccgcaatgtc cagcaacact
     3901 gggatggtgt cgtttcggcc attgcttaag ttcagaagtg ccttgggcag gcaggtcttc
     3961 cctgtggatg gttcccggaa ctcctcatca gtcaggcgct tcttgtgggt cagcaagtag
     4021 gagagcagtc cgtccaggtc ggcagtggag ccccgggaca cgatgtcaaa gaggatgggc
     4081 cggttgaaga ctttgaggat ggggggtggc tggggggcgg gagctttggg gctctgtggc
     4141 tgcttctcta cgaccttcct cctccatctc ttgttgtcac tggggtggtg ccggtaagtg
     4201 ccatagtcga acaacgaatc catgggcgct ttcttgggcc caggcactac tgaggactca
     4261 tacagggtgg actccagcag gtcaatgggg ttgggaaccc ccttgcggaa agcgccctgg
     4321 aacttcatac gcaggtttgg acgtccatcc ccggggccag cagggcggct agcatccact
     4381 ggtgaaagag aagaggagcc ttcctctccc tcaaacaggt tggccaggga agagaggggg
     4441 aaggcctccc caccagaagt gccactctcg tctccagggg gctcagccac atccccaggc
     4501 gctgcacggg ggccatcacc aggatctgcc atggtggcgg tacctctagc ataacttcgt
     4561 ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg ttattcttta
     4621 gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatga attcaaataa
     4681 aatacgaaat gcacgtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     4741 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4801 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     4861 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     4921 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4981 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5041 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5101 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     5161 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5221 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     5281 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5341 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     5401 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     5461 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5521 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5581 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5641 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5701 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5761 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5821 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5881 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5941 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6001 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6061 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6121 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6181 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6241 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6301 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6361 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6421 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6481 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6541 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6601 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6661 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6721 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6781 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6841 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6901 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     6961 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7021 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7081 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7141 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7201 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7261 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7321 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7381 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7441 tggccttttg ctcacatgt
//
