LOCUS       sequence-370168-        5727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of GCaMP8s under the control of GfaABC1D 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5727)
  TITLE     GCaMP8/jGCaMP8 AAV plasmids
REFERENCE   2  (bases 1 to 5727)
  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-cyto-GCaMP8s
FEATURES             Location/Qualifiers
     source          1..5727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          968..1100
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1083..1102
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1145..1163
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1180..1189
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1192..1209
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1491..1512
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1618..1639)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1879..1898)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2481..2602
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2492..2511)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2546..2565
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2764..2893
                     /label=AAV2 ITR
     primer_bind     complement(2912..2929)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2912..2928)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2921..2943)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3070..3525
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3157..3176)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3367..3388
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3551..3655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3656..3724,3725..4516)
                     /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      4687..5275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5176..5195
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5563..5584
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5599..5604,5605..5622,5623..5629)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5637..5653
                     /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     5642..5664
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5661..5677
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5661..5677
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   join(5715..5727,1..117)
                     /label=AAV2 ITR
ORIGIN
        1 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
       61 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt
      121 agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg aagatctaac
      181 atatcctggt gtggagtagg ggacgctgct ctgacagagg ctcgggggcc tgagctggct
      241 ctgtgagctg gggaggaggc agacagccag gccttgtctg caagcagacc tggcagcatt
      301 gggctggccg ccccccaggg cctcctcttc atgcccagtg aatgactcac cttggcacag
      361 acacaatgtt cggggtgggc acagtgcctg cttcccgccg caccccagcc cccctcaaat
      421 gccttccgag aagcccattg agcagggggc ttgcattgca ccccagcctg acagcctggc
      481 atcttgggat aaaagcagca cagcccccta ggggctgccc ttgctgtgtg gcgccaccgg
      541 cggtggagaa caaggctcta ttcagcctgt gcccaggaaa ggggatcagg ggatgcccag
      601 gcatggacag tgggtggcag ggggggagag gagggctgtc tgcttcccag aagtccaagg
      661 acacaaatgg gtgaggggag agctctcccc atagctgggc tgcggcccaa ccccaccccc
      721 tcaggctatg ccagggggtg ttgccagggg cacccgggca tcgccagtct agcccactcc
      781 ttcataaagc cctcgcatcc caggagcgag cagagccaga gcaggttgga gaggagacgc
      841 atcacctccg ctgctcgcaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      901 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac
      961 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
     1021 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
     1081 ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag
     1141 tacttaatac gactcactat aggctagcct cgagaattcg ccaccatggg ccatcatcac
     1201 catcatcaca cgcgtcgcaa gaagaccttc aaggaggtgg ccaccgccgt gaagatcatc
     1261 gccatgctga tgggcctgaa gatcaacgtc tatatcaagg ccgacaagca gaagaacggc
     1321 atcaaggcga acttccacat ccgccacaac atcgaggacg gcggcgtgca gctcgcctac
     1381 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1441 ctgagcgtgg agtccaaact ttcgaaagac cccaacgaga agcgcgatca catggtcctg
     1501 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt
     1561 accggaggga gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1621 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc
     1681 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1741 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     1801 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag
     1861 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1921 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1981 atcctggggc acaagctgga gtacaacctg ccggaccaac tgactgaaga gcagatcgca
     2041 gaatacaaag aggctttctc cctatttgac aaggacgggg atgggacaat aacaaccaag
     2101 gagctgggga cggtgatgcg gtctctcgga cacaacccca cagaagcaga gctgcaggac
     2161 atgatcaatg aagtagatgc cgacggtgac ggcacaatcg acttccctga gttcctgaca
     2221 atgatggcaa gaaaaatgaa atacagggac acggaagaag aaattagaga agcgttcggt
     2281 gtgtttgata aggatggcaa tggctacatc agtgcagcag agcttcgcca cgtgatgaca
     2341 aaccttggag agaagttaac agatgaagag gttgatgaaa tgatcaggga agcagacatc
     2401 gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc gaagtaagcg
     2461 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2521 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2581 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2641 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     2701 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     2761 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2821 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2881 agcgagcgcg cagccttaat taacctaatt cactggccgt cgttttacaa cgtcgtgact
     2941 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3001 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3061 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3121 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3181 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3241 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3301 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3361 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3421 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3481 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     3541 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3601 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3661 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3721 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3781 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3841 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3901 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3961 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4021 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4081 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4141 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4201 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4261 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4321 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4381 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4441 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4501 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4561 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4621 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4681 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4741 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4801 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4861 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4921 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4981 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5041 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5101 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5161 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5221 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5281 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5341 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5401 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     5461 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5521 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     5581 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5641 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca gatttaatta
     5701 aggccttaat taggctgcgc gctcgct
//
