LOCUS       sequence-349053-        5832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of membrane bound GCaMP8s 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-jGCaMP8s
FEATURES             Location/Qualifiers
     source          1..5832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          652..784
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     767..786
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        829..847
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             981..998
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1280..1301
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1407..1428)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1668..1687)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2270..2391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2281..2300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2335..2354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2553..2682
                     /label=AAV2 ITR
     primer_bind     complement(2701..2718)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2701..2717)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2710..2732)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2859..3314
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2946..2965)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3156..3177
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3340..3444
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3445..3513,3514..4305)
                     /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      4476..5064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4965..4984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5352..5373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5388..5393,5394..5411,5412..5418)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5426..5442
                     /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     5431..5453
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5450..5466
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5450..5466
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5504..5633
                     /label=AAV2 ITR
ORIGIN
        1 agggcctcct cttcatgccc agtgaatgac tcaccttggc acagacacaa tgttcggggt
       61 gggcacagtg cctgcttccc gccgcacccc agcccccctc aaatgccttc cgagaagccc
      121 attgagcagg gggcttgcat tgcaccccag cctgacagcc tggcatcttg ggataaaagc
      181 agcacagccc cctaggggct gcccttgctg tgtggcgcca ccggcggtgg agaacaaggc
      241 tctattcagc ctgtgcccag gaaaggggat caggggatgc ccaggcatgg acagtgggtg
      301 gcaggggggg agaggagggc tgtctgcttc ccagaagtcc aaggacacaa atgggtgagg
      361 ggagagctct ccccatagct gggctgcggc ccaaccccac cccctcaggc tatgccaggg
      421 ggtgttgcca ggggcacccg ggcatcgcca gtctagccca ctccttcata aagccctcgc
      481 atcccaggag cgagcagagc cagagcaggt tggagaggag acgcatcacc tccgctgctc
      541 gcaagcttta ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac
      601 acaacagtct cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc
      661 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
      721 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
      781 acaggtgtcc actcccagtt caattacagc tcttaaggct agagtactta atacgactca
      841 ctataggcta gcctcgagaa ttcaccatgg gctgtggctg cagctcaaac cctgaagatg
      901 actggatgga gaacattgac gtgtgtgaga actgccatta tcccccgccg aagcttcgaa
      961 tggatccacc cgccaccatg catcatcacc atcatcacac gcgtcgcaag aagaccttca
     1021 aggaggtggc caccgccgtg aagatcatcg ccatgctgat gggcctgaag atcaacgtct
     1081 atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttccacatc cgccacaaca
     1141 tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg
     1201 gccccgtgct gctgcccgac aaccactacc tgagcgtgga gtccaaactt tcgaaagacc
     1261 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1321 tcggcatgga cgagctgtac aagggcggta ccggagggag catggtgagc aagggcgagg
     1381 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1441 agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt
     1501 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1561 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1621 ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggcaact
     1681 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1741 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacctgc
     1801 cggaccaact gactgaagag cagatcgcag aatacaaaga ggctttctcc ctatttgaca
     1861 aggacgggga tgggacaata acaaccaagg agctggggac ggtgatgcgg tctctcggac
     1921 acaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg
     1981 gcacaatcga cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa tacagggaca
     2041 cggaagaaga aattagagaa gcgttcggtg tgtttgataa ggatggcaat ggctacatca
     2101 gtgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca gatgaagagg
     2161 ttgatgaaat gatcagggaa gcagacatcg atggggatgg tcaggtaaac tacgaagagt
     2221 ttgtacaaat gatgacagcg aagtaagcgg ccgcttcgag cagacatgat aagatacatt
     2281 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2341 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2401 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     2461 taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga
     2521 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     2581 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     2641 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt aacctaattc
     2701 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     2761 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     2821 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     2881 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2941 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3001 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3061 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3121 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3181 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3241 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3301 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3361 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3421 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3481 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3541 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3601 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3661 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3721 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3781 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3841 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3901 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3961 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4021 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4081 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4141 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4201 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4261 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4321 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4381 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4441 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4501 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4561 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4621 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4681 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4741 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4801 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4861 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4921 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4981 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5041 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5101 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5161 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5221 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     5281 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     5341 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     5401 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     5461 tatgaccatg attacgccag atttaattaa ggccttaatt aggctgcgcg ctcgctcgct
     5521 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     5581 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt
     5641 aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga tctaacatat
     5701 cctggtgtgg agtaggggac gctgctctga cagaggctcg ggggcctgag ctggctctgt
     5761 gagctgggga ggaggcagac agccaggcct tgtctgcaag cagacctggc agcattgggc
     5821 tggccgcccc cc
//
