LOCUS       sequence-329815-        6421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GCaMP6f in CaMKII+ cells with low miRNA-128 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6421)
  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 Dec 9;40(50):9576-9588. doi: 
            10.1523/JNEUROSCI.2570-19.2020. Epub 2020 Nov 6.
  PUBMED    33158963
REFERENCE   2  (bases 1 to 6421)
  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"; doi: 
            "10.1523/JNEUROSCI.2570-19.2020"; journalName: "J Neurosci"; date: 
            "2020-12-9- 9"; volume: "40"; issue: "50"; pages: "9576-9588"
COMMENT     Sequence Label: pAAV-CaMKII-GCaMP6f-4x128-mAGNET
FEATURES             Location/Qualifiers
     source          1..6421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     regulatory      1503..1512
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1509..1517,1518..1535,1536..1538,1539..1571,1572..1574,
                     1575..1598,1599..1625,1626..1682,1683..1688,1689..1958,
                     1959..1976,1977..2411,2412..2417,2418..2861)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1605..1614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1895..1916
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2022..2043)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2283..2302)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3008..3596
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3061..3081)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3479..3490)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3599..3615)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3600..3616)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3645..3766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3656..3675)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3710..3729
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4145..4733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4225..4244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4904..5695,5696..5764))
                     /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     5527..5546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5765..5869)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5895..6350)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6032..6053)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6244..6263
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatct gaattccatt atggccttag gtaggtcact tcatctccat ggggttcttc
      241 ttctgatttt ctagaaaatg agatgggggt gcagagagct tcctcagtga cctgcccagg
      301 gtcacatcag aaatgtcaga gctagaactt gaactcagat tactaatctt aaattccatg
      361 ccttgggggc atgcaagtac gatatacaga aggagtgaac tcattagggc agatgaccaa
      421 tgagtttagg aaagaagagt ccagggcagg gtacatctac accacccgcc cagccctggg
      481 tgagtccagc cacgttcacc tcattatagt tgcctctctc cagtcctacc ttgacgggaa
      541 gcacaagcag aaactgggac aggagcccca ggagaccaaa tcttcatggt ccctctggga
      601 ggatgggtgg ggagagctgt ggcagaggcc tcaggagggg ccctgctgct cagtggtgac
      661 agataggggt gagaaagcag acagagtcat tccgtcagca ttctgggtct gtttggtact
      721 tcttctcacg ctaaggtggc ggtgtgatat gcacaatggc taaaaagcag ggagagctgg
      781 aaagaaacaa ggacagagac agaggccaag tcaaccagac caattcccag aggaagcaaa
      841 gaaaccatta cagagactac aagggggaag ggaaggagag atgaattagc ttcccctgta
      901 aaccttagaa cccagctgtt gccagggcaa cggggcaata cctgtctctt cagaggagat
      961 gaagttgcca gggtaactac atcctgtctt tctcaaggac catcccagaa tgtggcaccc
     1021 actagccgtt accatagcaa ctgcctcttt gccccactta atcccatccc gtctgttaaa
     1081 agggccctat agttggaggt gggggaggta ggaagagcga tgatcacttg tggactaagt
     1141 ttgttcgcat ccccttctcc aaccccctca gtacatcacc ctgggggaac agggtccact
     1201 tgctcctggg cccacacagt cctgcagtat tgtgtatata aggccagggc aaagaggagc
     1261 aggttttaaa gtgaaaggca ggcaggtgtt ggggaggcag ttaccggggc aacgggaaca
     1321 gggcgtttcg gaggtggttg ccatggggac ctggatgctg acgaaggctc gcgaggctgt
     1381 gagcagccac agtgccctgc tcagaagccc caagctcgtc agtcaagccg gttctccgtt
     1441 tgcactcagg agcacgggca ggcgagtggc ccctagttct gggggcagcg gggcgcgccg
     1501 ccgccaccat gggttctcat catcatcatc atcatggtat ggctagcatg actggtggac
     1561 agcaaatggg tcgggatctg tacgacgatg acgataagga tctcgccacc atggtcgact
     1621 catcacgtcg taagtggaat aagacaggtc acgcagtcag agctataggt cggctgagct
     1681 cactcgagaa cgtctatatc aaggccgaca agcagaagaa cggcatcaag gcgaacttca
     1741 agatccgcca caacatcgag gacggcggcg tacagctcgc ctaccactac cagcagaaca
     1801 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc gtgcagtcca
     1861 aactttcgaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1921 ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatgg
     1981 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2041 acgtaaacgg ccacaagttc agcgtgtccg gcgagggtga gggcgatgcc acctacggca
     2101 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2161 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2221 acgacttctt caagtccgcc atgcccgaag gctacatcca ggagcgcacc atcttcttca
     2281 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2341 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2401 tggagtacaa cctgccggac caactgactg aagagcagat cgcagaattt aaagaggaat
     2461 tctccctatt tgacaaggac ggggatggga caataacaac caaggagctg gggacggtga
     2521 tgcggtctct ggggcagaac cccacagaag cagagctgca ggacatgatc aatgaagtag
     2581 atgccgacgg tgacggcaca atcgacttcc ctgagttcct gacaatgatg gcaagaaaaa
     2641 tgaaatacag ggacacggaa gaagaaatta gagaagcgtt cggtgtgttt gataaggatg
     2701 gcaatggcta catcagtgca gcagagcttc gccacgtgat gacaaacctt ggagagaagt
     2761 taacagatga agaggttgat gaaatgatca gggaagcaga catcgatggg gatggtcagg
     2821 taaactacga agagtttgta caaatgatga cagcgaagtg aactagtgcg gccttaaaga
     2881 gaccggttca ctgtgacagt aaaagagacc ggttcactgt gagaatgaaa gagaccggtt
     2941 cactgtgatc ggaaaagaga ccggttcact gtgagcggcc gccctgcagg catgcaagct
     3001 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3061 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3121 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3181 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3241 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3301 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3361 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3421 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3481 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3541 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3601 ataccgtcga cctcgacccg ggcggccgct tcgagcagac atgataagat acattgatga
     3661 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3721 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3781 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     3841 cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt
     3901 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct
     3961 ctctgcgcgc tcgctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4021 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4081 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4141 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4201 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4261 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4321 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4381 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4441 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4501 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4561 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4621 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4681 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4741 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4801 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4861 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4921 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4981 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5041 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5101 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5161 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5221 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5281 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5341 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5401 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5461 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5521 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5581 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5641 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5701 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5761 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5821 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5881 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5941 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     6001 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     6061 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     6121 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     6181 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     6241 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     6301 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     6361 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6421 g
//
