LOCUS       sequence-201051-        5302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of membrane bound EGFP from GfaABC1D promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5302)
  AUTHORS   Shigetomi E, Bushong EA, Haustein MD, Tong X, Jackson-Weaver O, 
            Kracun S, Xu J, Sofroniew MV, Ellisman MH, Khakh BS
  TITLE     Imaging calcium microdomains within entire astrocyte territories and
            endfeet with GCaMPs expressed using adeno-associated viruses.
  JOURNAL   J Gen Physiol. 2013 Apr 15.
  PUBMED    23589582
REFERENCE   2  (bases 1 to 5302)
  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"; journalName: "J Gen 
            Physiol. 2013 Apr 15."
COMMENT     Sequence Label: pAAV.GfaABC1D.PI.Lck-GFP.SV40
FEATURES             Location/Qualifiers
     source          1..5302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1158..1176
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1304..1313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1310..1312,1313..1315,1316..2029)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1310..1312,1313..1315,1316..2026)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1355..1376)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1616..1635)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1963..1984
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2069..2190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2080..2099)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2134..2153
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2352..2481
                     /label=AAV2 ITR
     primer_bind     complement(2500..2517)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2500..2516)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2509..2531)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2658..3113
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2745..2764)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2955..2976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3139..3243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3244..3312,3313..4104)
                     /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      4275..4863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4764..4783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5151..5172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5187..5192,5193..5210,5211..5217)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5225..5241
                     /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     5230..5252
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5249..5265
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5249..5265
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatct aacatatcct ggtgtggagt aggggacgct gctctgacag aggctcgggg
      241 gcctgagctg gctctgtgag ctggggagga ggcagacagc caggccttgt ctgcaagcag
      301 acctggcagc attgggctgg ccgcccccca gggcctcctc ttcatgccca gtgaatgact
      361 caccttggca cagacacaat gttcggggtg ggcacagtgc ctgcttcccg ccgcacccca
      421 gcccccctca aatgccttcc gagaagccca ttgagcaggg ggcttgcatt gcaccccagc
      481 ctgacagcct ggcatcttgg gataaaagca gcacagcccc ctaggggctg cccttgctgt
      541 gtggcgccac cggcggtgga gaacaaggct ctattcagcc tgtgcccagg aaaggggatc
      601 aggggatgcc caggcatgga cagtgggtgg caggggggga gaggagggct gtctgcttcc
      661 cagaagtcca aggacacaaa tgggtgaggg gagagctctc cccatagctg ggctgcggcc
      721 caaccccacc ccctcaggct atgccagggg gtgttgccag gggcacccgg gcatcgccag
      781 tctagcccac tccttcataa agccctcgca tcccaggagc gagcagagcc agagcaggtt
      841 ggagaggaga cgcatcacct ccgctgctcg caagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtacttaa tacgactcac tataggctag cctcgagaat tcaccatggg
     1201 ctgtggctgc agctcaaacc ctgaagatga ctggatggag aacattgacg tgtgtgagaa
     1261 ctgccattat cccccgccga agcttcgaat tgatccaccg gtcgccacca tggtgagcaa
     1321 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1381 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1441 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1501 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1561 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1621 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1681 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1741 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1801 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1861 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1921 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1981 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat ctagagtcga
     2041 cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     2101 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2161 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2221 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2281 aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat
     2341 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     2401 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     2461 agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg
     2521 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     2581 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     2641 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2701 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2761 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2821 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2881 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2941 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3001 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3061 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3121 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3181 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3241 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3301 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3361 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3421 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3481 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3541 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3601 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3661 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3721 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3781 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3841 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3901 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3961 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4021 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4081 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4141 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4201 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4261 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4321 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4381 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4441 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4501 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4561 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4621 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4681 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4741 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4801 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4861 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4921 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4981 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5041 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5101 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     5161 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     5221 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaga
     5281 tttaattaag gccttaatta gg
//
