LOCUS       sequence-349055-        5832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of membrane bound GCaMP8m under the control 
            of GfaABC1D promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  TITLE     Looger Lab GCaMP8 plasmids
REFERENCE   2  (bases 1 to 5832)
  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: pZac2.1-GfaABC1D-lck-jGCaMP8m
FEATURES             Location/Qualifiers
     source          1..5832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          812..944
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     927..946
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        989..1007
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1141..1158
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1440..1461
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1567..1588)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1828..1847)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2430..2551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2441..2460)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2495..2514
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2713..2842
                     /label=AAV2 ITR
     primer_bind     complement(2861..2878)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2861..2877)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2870..2892)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3019..3474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3106..3125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3316..3337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3500..3604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3605..3673,3674..4465)
                     /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"
     rep_origin      4636..5224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5125..5144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5512..5533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5548..5553,5554..5571,5572..5578)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5586..5602
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     5591..5613
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5610..5626
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5610..5626
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5664..5793
                     /label=AAV2 ITR
ORIGIN
        1 agccatgctc taggaagatc taacatatcc tggtgtggag taggggacgc tgctctgaca
       61 gaggctcggg ggcctgagct ggctctgtga gctggggagg aggcagacag ccaggccttg
      121 tctgcaagca gacctggcag cattgggctg gccgcccccc agggcctcct cttcatgccc
      181 agtgaatgac tcaccttggc acagacacaa tgttcggggt gggcacagtg cctgcttccc
      241 gccgcacccc agcccccctc aaatgccttc cgagaagccc attgagcagg gggcttgcat
      301 tgcaccccag cctgacagcc tggcatcttg ggataaaagc agcacagccc cctaggggct
      361 gcccttgctg tgtggcgcca ccggcggtgg agaacaaggc tctattcagc ctgtgcccag
      421 gaaaggggat caggggatgc ccaggcatgg acagtgggtg gcaggggggg agaggagggc
      481 tgtctgcttc ccagaagtcc aaggacacaa atgggtgagg ggagagctct ccccatagct
      541 gggctgcggc ccaaccccac cccctcaggc tatgccaggg ggtgttgcca ggggcacccg
      601 ggcatcgcca gtctagccca ctccttcata aagccctcgc atcccaggag cgagcagagc
      661 cagagcaggt tggagaggag acgcatcacc tccgctgctc gcaagcttta ttgcggtagt
      721 ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct cgaacttaag
      781 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
      841 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      901 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt
      961 caattacagc tcttaaggct agagtactta atacgactca ctataggcta gcctcgagaa
     1021 ttcaccatgg gctgtggctg cagctcaaac cctgaagatg actggatgga gaacattgac
     1081 gtgtgtgaga actgccatta tcccccgccg aagcttcgaa tggatccacc cgccaccatg
     1141 catcatcacc atcatcacac gcgtcgcaag aagaccttca aggaggtggc caccgccgtg
     1201 aagatcatcg gccgcctgat gggcctgaag atcaacgtct atatcaaggc cgacaagcag
     1261 aagaacggca tcaaggcgaa cttccacatc cgccacaaca tcgaggacgg cggcgtgcag
     1321 ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1381 aaccactacc tgagcgtgga gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac
     1441 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1501 aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg
     1561 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1621 ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1681 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1741 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1801 atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1861 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1921 gacggcaaca tcctggggca caagctggag tacaacctgc cggaccaact gactgaagag
     1981 cagatcgcag aatacaaaga ggctttctcc ctatttgaca aggacgggga tgggacaata
     2041 acaaccaagg agctggggac ggtgatgcgg tctctggggc agaaccccac agaagcagag
     2101 ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacaatcga cttccctgag
     2161 ttcctgacaa tgatggcaag aaaaatgaaa tacagggaca cggaagaaga aattagagaa
     2221 gcgttcggtg tgtttgataa ggatggcaat ggctacatca gtgcagcaga gcttcgccac
     2281 gtgatgacaa accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggaa
     2341 gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg
     2401 aagtaagcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2461 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2521 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2581 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     2641 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     2701 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2761 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2821 cagtgagcga gcgagcgcgc agccttaatt aacctaattc actggccgtc gttttacaac
     2881 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     2941 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3001 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3061 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3121 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3181 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3241 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3301 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3361 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3421 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     3481 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3541 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3601 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3661 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3721 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3781 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3841 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3901 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3961 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4021 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4081 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4141 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4201 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4261 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4321 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4381 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4441 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4501 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4561 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4621 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4681 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4741 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4801 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4861 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4921 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4981 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5041 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5101 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5161 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5221 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5281 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5341 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5401 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     5461 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     5521 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     5581 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag
     5641 atttaattaa ggccttaatt aggctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     5701 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     5761 agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct
     5821 acttatctac gt
//
