LOCUS       sequence-349032-        5926 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV-mediated expression of histone fused GCaMP8m under the synapsin 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5926)
  TITLE     Looger Lab GCaMP8 plasmids
REFERENCE   2  (bases 1 to 5926)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AAV-Syn-H2B-jGCaMP8s-WPRE
FEATURES             Location/Qualifiers
     source          1..5926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(112..700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(192..211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(871..1662,1663..1731))
                     /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     1494..1513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1732..1836)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1862..2317)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1999..2020)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2211..2230
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2389..2518
                     /label=AAV2 ITR
     promoter        2600..3047
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             3081..3458
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             3474..3491
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     3773..3794
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3900..3921)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4161..4180)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4752..5340
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4805..4825)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5223..5234)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5343..5359)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5344..5360)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5389..5510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5400..5419)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5454..5473
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5672..5801
                     /label=AAV2 ITR
ORIGIN
        1 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
       61 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      121 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      181 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      241 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      301 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      361 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      421 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      481 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      541 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      601 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      661 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      721 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
      781 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
      841 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
      901 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
      961 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     1021 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1081 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1141 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1201 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1261 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1321 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1381 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1441 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     1501 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1561 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     1621 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     1681 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     1741 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1801 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     1861 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     1921 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1981 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2041 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2101 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2161 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2221 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2281 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     2341 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc
     2401 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc
     2461 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg
     2521 tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag gaagatctct
     2581 gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg
     2641 gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa
     2701 ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac
     2761 tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc
     2821 gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg
     2881 gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg
     2941 agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc
     3001 agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg aattcaagct
     3061 gctagcaagg atccgccacc atgccagagc cagcgaagtc tgctcccgcc ccgaaaaagg
     3121 gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc
     3181 gcaaggagag ctattccatc tatgtgtaca aggttctgaa gcaggtccac cctgacaccg
     3241 gcatttcgtc caaggccatg ggcatcatga attcgtttgt gaacgacatt ttcgagcgca
     3301 tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc acctccaggg
     3361 agatccagac ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg
     3421 agggtactaa ggccatcacc aagtacacca gcgctaagga cccaccggtc atgcatcatc
     3481 accatcatca cacgcgtcgc aagaagacct tcaaggaggt ggccaccgcc gtgaagatca
     3541 tcgccatgct gatgggcctg aagatcaacg tctatatcaa ggccgacaag cagaagaacg
     3601 gcatcaaggc gaacttccac atccgccaca acatcgagga cggcggcgtg cagctcgcct
     3661 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3721 acctgagcgt ggagtccaaa ctttcgaaag accccaacga gaagcgcgat cacatggtcc
     3781 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg
     3841 gtaccggagg gagcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     3901 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggtgagg
     3961 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     4021 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     4081 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg
     4141 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     4201 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     4261 acatcctggg gcacaagctg gagtacaacc tgccggacca actgactgaa gagcagatcg
     4321 cagaatacaa agaggctttc tccctatttg acaaggacgg ggatgggaca ataacaacca
     4381 aggagctggg gacggtgatg cggtctctcg gacacaaccc cacagaagca gagctgcagg
     4441 acatgatcaa tgaagtagat gccgacggtg acggcacaat cgacttccct gagttcctga
     4501 caatgatggc aagaaaaatg aaatacaggg acacggaaga agaaattaga gaagcgttcg
     4561 gtgtgtttga taaggatggc aatggctaca tcagtgcagc agagcttcgc cacgtgatga
     4621 caaaccttgg agagaagtta acagatgaag aggttgatga aatgatcagg gaagcagaca
     4681 tcgatgggga tggtcaggta aactacgaag agtttgtaca aatgatgaca gcgaagtaaa
     4741 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     4801 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     4861 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     4921 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     4981 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5041 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     5101 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     5161 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     5221 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     5281 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     5341 atcgataccg tcgacctcga cccgggcggc cgcttcgagc agacatgata agatacattg
     5401 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5461 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     5521 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     5581 aaaacctcta caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat
     5641 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     5701 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     5761 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc
     5821 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5881 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactc
//
