LOCUS       sequence-215846-        5780 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV vector that targets expression to GABAergic inhibitory neurons 
            in rodent cortex with 98% selectivity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5780)
  AUTHORS   Keaveney MK, Tseng HA, Ta TL, Gritton HJ, Man HY, Han X
  TITLE     A MicroRNA-Based Gene-Targeting Tool for Virally Labeling 
            Interneurons in the Rodent Cortex.
  JOURNAL   Cell Rep. 2018 Jul 10;24(2):294-303. doi: 
            10.1016/j.celrep.2018.06.049.
  PUBMED    29996091
REFERENCE   2  (bases 1 to 5780)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2018.06"; journalName: "Cell Rep"; date: 
            "2018-07-10- 10"; volume: "24"; issue: "2"; pages: "294-303"
COMMENT     Sequence Label: AAV-hSyn-EGFP-8X2C (AAV GABA mAGNET)
FEATURES             Location/Qualifiers
     source          1..5780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        224..671
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(693..695,696..698,699..1412)
                     /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(693..695,696..698,699..1409)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(738..759)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(999..1018)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1346..1367
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1910..2498
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1963..1983)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2381..2392)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2501..2517)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2502..2518)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2547..2668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2558..2577)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2612..2631
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2830..2959
                     /label=AAV2 ITR
     primer_bind     complement(2978..2995)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2978..2994)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2987..3009)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3136..3591
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3223..3242)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3433..3454
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3617..3721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3722..3790,3791..4582)
                     /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"
     primer_bind     complement(3940..3959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4753..5341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5242..5261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5629..5650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5665..5670,5671..5688,5689..5695)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5703..5719
                     /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     5708..5730
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5727..5743
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5727..5743
                     /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 gaattctcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
      241 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      301 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      361 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      421 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      481 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      541 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      601 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      661 tgagagcgca gtcgagagga tccgccgcca ccatggtgag caagggcgag gagctgttca
      721 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      781 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatttgca
      841 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      901 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      961 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1021 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1081 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1141 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1201 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1261 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1321 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1381 tcactctcgg catggacgag ctgtacaagt aaactagtgc ggccttaaag agaccggttc
     1441 actgtgacag taaaagagac cggttcactg tgagaatgaa agagaccggt tcactgtgat
     1501 cggaaaagag accggttcac tgtgagcggc cttaaagaga ccggttcact gtgacagtaa
     1561 aagagaccgg ttcactgtga gaatgaaaga gaccggttca ctgtgatcgg aaaagagacc
     1621 ggttcactgt gagcggcctt gaaacccagc agacaatgta gctcagtaga aacccagcag
     1681 acaatgtagc tgaatggaaa cccagcagac aatgtagctt cggagaaacc cagcagacaa
     1741 tgtagctgcg gccttgaaac ccagcagaca atgtagctca gtagaaaccc agcagacaat
     1801 gtagctgaat ggaaacccag cagacaatgt agcttcggag aaacccagca gacaatgtag
     1861 ctgcggccgc gactctagag tcgacctgca ggcatgcaag cttatcgata atcaacctct
     1921 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1981 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2041 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2101 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2161 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2221 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2281 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     2341 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2401 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2461 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc
     2521 cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2581 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2641 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2701 ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     2761 aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca
     2821 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     2881 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     2941 tgagcgagcg agcgcgcagc cttaattaac ctaattcact ggccgtcgtt ttacaacgtc
     3001 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     3061 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     3121 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3181 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3241 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3301 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3361 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3421 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3481 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3541 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     3601 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3661 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3721 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3781 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3841 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3901 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3961 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4021 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4081 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4141 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4201 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4261 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4321 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4381 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4441 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4501 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4561 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4621 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4681 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4741 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4801 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4861 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4921 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4981 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5041 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5101 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5161 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5221 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5281 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5341 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5401 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5461 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5521 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     5581 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     5641 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     5701 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccagatt
     5761 taattaaggc cttaattagg
//
