LOCUS       sequence-202337-        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"
     protein_bind    96..117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(132..137,138..155,156..162)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    170..186
                     /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     175..197
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     194..210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     194..210
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   248..377
                     /label=AAV2 ITR
     intron          1228..1360
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1343..1362
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1405..1423
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1551..1560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1557..1559,1560..1562,1563..2276)
                     /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(1557..1559,1560..1562,1563..2273)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1602..1623)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1863..1882)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2210..2231
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2316..2437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2327..2346)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2381..2400
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2599..2728
                     /label=AAV2 ITR
     primer_bind     complement(2747..2764)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2747..2763)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2756..2778)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2905..3360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2992..3011)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3202..3223
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3386..3490
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3491..3559,3560..4351)
                     /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      4522..5110
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5011..5030
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
       61 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
      121 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
      181 taacaatttc acacaggaaa cagctatgac catgattacg ccagatttaa ttaaggcctt
      241 aattaggctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
      301 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
      361 catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat ctacgtagcc
      421 atgctctagg aagatctaac atatcctggt gtggagtagg ggacgctgct ctgacagagg
      481 ctcgggggcc tgagctggct ctgtgagctg gggaggaggc agacagccag gccttgtctg
      541 caagcagacc tggcagcatt gggctggccg ccccccaggg cctcctcttc atgcccagtg
      601 aatgactcac cttggcacag acacaatgtt cggggtgggc acagtgcctg cttcccgccg
      661 caccccagcc cccctcaaat gccttccgag aagcccattg agcagggggc ttgcattgca
      721 ccccagcctg acagcctggc atcttgggat aaaagcagca cagcccccta ggggctgccc
      781 ttgctgtgtg gcgccaccgg cggtggagaa caaggctcta ttcagcctgt gcccaggaaa
      841 ggggatcagg ggatgcccag gcatggacag tgggtggcag ggggggagag gagggctgtc
      901 tgcttcccag aagtccaagg acacaaatgg gtgaggggag agctctcccc atagctgggc
      961 tgcggcccaa ccccaccccc tcaggctatg ccagggggtg ttgccagggg cacccgggca
     1021 tcgccagtct agcccactcc ttcataaagc cctcgcatcc caggagcgag cagagccaga
     1081 gcaggttgga gaggagacgc atcacctccg ctgctcgcaa gctttattgc ggtagtttat
     1141 cacagttaaa ttgctaacgc agtcagtgct tctgacacaa cagtctcgaa cttaagctgc
     1201 agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga
     1261 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     1321 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat
     1381 tacagctctt aaggctagag tacttaatac gactcactat aggctagcct cgagaattca
     1441 ccatgggctg tggctgcagc tcaaaccctg aagatgactg gatggagaac attgacgtgt
     1501 gtgagaactg ccattatccc ccgccgaagc ttcgaattga tccaccggtc gccaccatgg
     1561 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1621 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1681 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1741 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1801 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1861 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1921 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1981 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2041 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2101 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2161 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2221 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatcta
     2281 gagtcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     2341 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2401 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2461 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     2521 atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg
     2581 ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     2641 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     2701 cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc taattcactg gccgtcgttt
     2761 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     2821 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2881 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2941 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3001 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3061 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3121 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3181 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3241 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3301 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     3361 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3421 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3481 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3541 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3601 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3661 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3721 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3781 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3841 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3901 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3961 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4021 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4081 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4141 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4201 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4261 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4321 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4381 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4441 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4501 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4561 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4621 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4681 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4741 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4801 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4861 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4921 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4981 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5041 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5101 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5161 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5221 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5281 ggaagcggaa gagcgcccaa ta
//
