LOCUS       sequence-349056-        5832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of membrane bound GCaMP8f 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-jGCaMP8f
FEATURES             Location/Qualifiers
     source          1..5832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          782..914
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     897..916
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        959..977
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1111..1128
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1410..1431
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1537..1558)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1798..1817)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2400..2521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2411..2430)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2465..2484
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2683..2812
                     /label=AAV2 ITR
     primer_bind     complement(2831..2848)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2831..2847)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2840..2862)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2989..3444
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3076..3095)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3286..3307
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3470..3574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3575..3643,3644..4435)
                     /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      4606..5194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5095..5114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5482..5503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5518..5523,5524..5541,5542..5548)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5556..5572
                     /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     5561..5583
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5580..5596
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5580..5596
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5634..5763
                     /label=AAV2 ITR
ORIGIN
        1 tggtgtggag taggggacgc tgctctgaca gaggctcggg ggcctgagct ggctctgtga
       61 gctggggagg aggcagacag ccaggccttg tctgcaagca gacctggcag cattgggctg
      121 gccgcccccc agggcctcct cttcatgccc agtgaatgac tcaccttggc acagacacaa
      181 tgttcggggt gggcacagtg cctgcttccc gccgcacccc agcccccctc aaatgccttc
      241 cgagaagccc attgagcagg gggcttgcat tgcaccccag cctgacagcc tggcatcttg
      301 ggataaaagc agcacagccc cctaggggct gcccttgctg tgtggcgcca ccggcggtgg
      361 agaacaaggc tctattcagc ctgtgcccag gaaaggggat caggggatgc ccaggcatgg
      421 acagtgggtg gcaggggggg agaggagggc tgtctgcttc ccagaagtcc aaggacacaa
      481 atgggtgagg ggagagctct ccccatagct gggctgcggc ccaaccccac cccctcaggc
      541 tatgccaggg ggtgttgcca ggggcacccg ggcatcgcca gtctagccca ctccttcata
      601 aagccctcgc atcccaggag cgagcagagc cagagcaggt tggagaggag acgcatcacc
      661 tccgctgctc gcaagcttta ttgcggtagt ttatcacagt taaattgcta acgcagtcag
      721 tgcttctgac acaacagtct cgaacttaag ctgcagaagt tggtcgtgag gcactgggca
      781 ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag
      841 acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc
      901 ctttctctcc acaggtgtcc actcccagtt caattacagc tcttaaggct agagtactta
      961 atacgactca ctataggcta gcctcgagaa ttcaccatgg gctgtggctg cagctcaaac
     1021 cctgaagatg actggatgga gaacattgac gtgtgtgaga actgccatta tcccccgccg
     1081 aagcttcgaa tggatccacc cgccaccatg catcatcacc atcatcacac gcgtcgcaag
     1141 aagaccttca aggaggtggc caccgccgtg aagatcatcg cccgcctgat gggcctgaag
     1201 atcaacgtct atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttccacatc
     1261 cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc
     1321 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgga gtccaaactt
     1381 tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1441 gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag catggtgagc
     1501 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1561 aacggccaca agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg
     1621 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1681 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1741 ttcttcaagt ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac
     1801 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1861 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1921 tacaacctgc cggaccaact gactgaagag cagatcgcag aatttaaaga ggctttctcc
     1981 ctatttgaca aggacgggga tgggacaata acaaccaagg agctggggac ggtgatgcgg
     2041 tctctcggac tgaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc
     2101 gacggtgacg gcacaatcga cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa
     2161 tacagggaca cggaagaaga aattagagaa gcgttcggtg tgtttgataa ggatggcaat
     2221 ggctacatca gtgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca
     2281 gatgaagagg ttgatgaaat gatcagggaa gcagacatcg atggggatgg tcaggtaaac
     2341 tacgaagagt ttgtacaaat gatgacagcg aagtaagcgg ccgcttcgag cagacatgat
     2401 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2461 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2521 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     2581 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg
     2641 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     2701 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     2761 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt
     2821 aacctaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     2881 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     2941 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     3001 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3061 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3121 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3181 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3241 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3301 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3361 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3421 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3481 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3541 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3601 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3661 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3721 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3781 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3841 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3901 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3961 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4021 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4081 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4141 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4201 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4261 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4321 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4381 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4441 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4501 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4561 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4621 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4681 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4741 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4801 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4861 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4921 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4981 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5041 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5101 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5161 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5221 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5281 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5341 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5401 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5461 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     5521 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5581 aggaaacagc tatgaccatg attacgccag atttaattaa ggccttaatt aggctgcgcg
     5641 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     5701 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     5761 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
     5821 tctaacatat cc
//
