LOCUS       62056_206        6878 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector AAV-CaMKII-GCaMP6f-4X128-mAGNET, complete sequence.
ACCESSION   MW139901
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6878)
  AUTHORS   Keaveney MK, Rahsepar B, Tseng HA, Fernandez FR, Mount RA, Ta T, 
            White JA, Berg J, Han X.
  TITLE     CamKIIalpha Positive Interneurons Identified via A microRNA Based 
            Viral Gene Targeting Strategy
  JOURNAL   J Neurosci (2020) In press
  PUBMED    33158963
REFERENCE   2  (bases 1 to 6878)
  AUTHORS   Keaveney MK, Rahsepar B, Tseng HA, Fernandez FR, Mount RA, Ta T, 
            White JA, Berg J, Han X.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-OCT-2020) Biomedical Engineering, Boston University, 
            44 Cummington Mall, Boston, MA 02215, USA
REFERENCE   3  (bases 1 to 6878)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Neurosci 
            (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-OCT-2020) Biomedical Engineering, Boston University, 44 
            Cummington Mall, Boston, MA 02215, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    228..249
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        264..294
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /label=lac operator
                     /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)."
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   380..480
     promoter        576..1868
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             1888..3237
                     /label=GCaMP6f
                     /note="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
     misc_binding    3255..3275
                     /label=miRNA-128 binding site
                     /bound_moiety="miRNA-128"
     misc_binding    3281..3301
                     /label=miRNA-128 binding site
                     /bound_moiety="miRNA-128"
     misc_binding    3307..3327
                     /label=miRNA-128 binding site
                     /bound_moiety="miRNA-128"
     misc_binding    3333..3353
                     /label=miRNA-128 binding site
                     /bound_moiety="miRNA-128"
     misc_feature    3387..3975
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(3978..3994)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(4024..4145)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   4248..4437
     primer_bind     complement(4455..4471)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4613..5068
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5094..5198
                     /label=AmpR promoter
     CDS             5199..6056
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6230..6818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      361 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      421 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      481 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      541 atctacgtag ccatgctcta ggaagatctg aattccatta tggccttagg tcaggtactt
      601 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt
      661 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt
      721 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact
      781 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca
      841 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc
      901 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat
      961 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc
     1021 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat
     1081 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct
     1141 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc
     1201 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga
     1261 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac
     1321 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc
     1381 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa
     1441 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat
     1501 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc
     1561 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa
     1621 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt
     1681 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga
     1741 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca
     1801 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg
     1861 ggggcagcgg ggcgcgccgc cgccaccatg ggttctcatc atcatcatca tcatggtatg
     1921 gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat
     1981 ctcgccacca tggtcgactc atcacgtcgt aagtggaata agacaggtca cgcagtcaga
     2041 gctataggtc ggctgagctc actcgagaac gtctatatca aggccgacaa gcagaagaac
     2101 ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     2161 taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2221 tacctgagcg tgcagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc
     2281 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     2341 ggtaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2401 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag
     2461 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2521 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     2581 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag
     2641 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2701 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2761 aacatcctgg ggcacaagct ggagtacaac ctgccggacc aactgactga agagcagatc
     2821 gcagaattta aagaggaatt ctccctattt gacaaggacg gggatgggac aataacaacc
     2881 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag
     2941 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
     3001 acaatgatgg caagaaaaat gaaatacagg gacacggaag aagaaattag agaagcgttc
     3061 ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
     3121 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
     3181 atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagtga
     3241 actagtgcgg ccttaaagag accggttcac tgtgacagta aaagagaccg gttcactgtg
     3301 agaatgaaag agaccggttc actgtgatcg gaaaagagac cggttcactg tgagcggccg
     3361 ccctgcaggc atgcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3421 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3481 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3541 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3601 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3661 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3721 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3781 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     3841 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3901 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3961 gggccgcctc cccgcatcga taccgtcgac ctcgacccgg gcggccgctt cgagcagaca
     4021 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     4081 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     4141 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     4201 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag
     4261 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     4321 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4381 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt
     4441 aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     4501 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     4561 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     4621 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4681 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4741 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4801 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4861 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4921 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4981 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5041 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     5101 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5161 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5221 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5281 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5341 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5401 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5461 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5521 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5581 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5641 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5701 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5761 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5821 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5881 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5941 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6001 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6061 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6121 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6181 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6241 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6301 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6361 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6421 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6481 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6541 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6601 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6661 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6721 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6781 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6841 tacggttcct ggccttttgc tggccttttg ctcacatg
//
