LOCUS       sequence-349048-        6333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of GCaMP8s under the EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6333)
  TITLE     Looger Lab GCaMP8 plasmids
REFERENCE   2  (bases 1 to 6333)
  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"
COMMENT     Sequence Label: AAV-EF1a-jGCaMP8s-WPRE
FEATURES             Location/Qualifiers
     source          1..6333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     promoter        280..1457
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          511..1449
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1406..1426
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1496..1513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1795..1816
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1922..1943)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2183..2202)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2774..3362
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2827..2847)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3245..3256)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3365..3381)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3366..3382)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3411..3532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3422..3441)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3476..3495
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3694..3823
                     /label=AAV2 ITR
     rep_origin      complement(4060..4648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4140..4159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4819..5610,5611..5679))
                     /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     5442..5461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5680..5784)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5810..6265)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5947..5968)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6159..6178
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggaaga tccaagcttt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
      241 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
      301 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
      361 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
      421 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
      481 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
      541 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
      601 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
      661 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
      721 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
      781 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
      841 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
      901 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
      961 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     1021 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     1081 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     1141 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     1201 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     1261 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     1321 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     1381 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     1441 ccatttcagg tgtcgtggaa ttcaagctgc tagcaaggat ccacccgcca ccatgcatca
     1501 tcaccatcat cacacgcgtc gcaagaagac cttcaaggag gtggccaccg ccgtgaagat
     1561 catcgccatg ctgatgggcc tgaagatcaa cgtctatatc aaggccgaca agcagaagaa
     1621 cggcatcaag gcgaacttcc acatccgcca caacatcgag gacggcggcg tgcagctcgc
     1681 ctaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1741 ctacctgagc gtggagtcca aactttcgaa agaccccaac gagaagcgcg atcacatggt
     1801 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     1861 cggtaccgga gggagcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1921 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggtga
     1981 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2041 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     2101 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacatcca
     2161 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2221 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2281 caacatcctg gggcacaagc tggagtacaa cctgccggac caactgactg aagagcagat
     2341 cgcagaatac aaagaggctt tctccctatt tgacaaggac ggggatggga caataacaac
     2401 caaggagctg gggacggtga tgcggtctct cggacacaac cccacagaag cagagctgca
     2461 ggacatgatc aatgaagtag atgccgacgg tgacggcaca atcgacttcc ctgagttcct
     2521 gacaatgatg gcaagaaaaa tgaaatacag ggacacggaa gaagaaatta gagaagcgtt
     2581 cggtgtgttt gataaggatg gcaatggcta catcagtgca gcagagcttc gccacgtgat
     2641 gacaaacctt ggagagaagt taacagatga agaggttgat gaaatgatca gggaagcaga
     2701 catcgatggg gatggtcagg taaactacga agagtttgta caaatgatga cagcgaagta
     2761 aaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2821 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2881 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2941 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3001 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3061 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3121 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3181 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3241 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3301 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3361 gcatcgatac cgtcgacctc gacccgggcg gccgcttcga gcagacatga taagatacat
     3421 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3481 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3541 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa
     3601 gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat ggctacgtag
     3661 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     3721 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3781 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg
     3841 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3901 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4501 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4741 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4801 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4861 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4921 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4981 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5041 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5101 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5161 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5221 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5281 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5341 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5401 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5461 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5521 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5581 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5641 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5701 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5761 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     5821 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5881 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     5941 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     6001 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     6061 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     6121 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     6181 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     6241 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     6301 gggcgatcgg tgcgggcctc ttcgctatta cgc
//
