LOCUS       sequence-226454-        7153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a GABA transporter GAT-3 (Slc6a11).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7153)
  AUTHORS   Yu X, Taylor AMW, Nagai J, Golshani P, Evans CJ, Coppola G, Khakh BS
  TITLE     Reducing Astrocyte Calcium Signaling In Vivo Alters Striatal 
            Microcircuits and Causes Repetitive Behavior.
  JOURNAL   Neuron. 2018 Aug 27. pii: S0896-6273(18)30688-3. doi: 
            10.1016/j.neuron.2018.08.015.
  PUBMED    30174118
REFERENCE   2  (bases 1 to 7153)
  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: "Neuron. 
            2018 Aug 27. pii: S0896-6273(18)30688-3. doi: 
            10.1016/j.neuron.2018.08.015."
COMMENT     Sequence Label: pZac2.1-GfaABC1D-GAT3-HA-mCherry
FEATURES             Location/Qualifiers
     source          1..7153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    40..161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(51..70)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     105..124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   323..452
                     /label=AAV2 ITR
     primer_bind     complement(471..488)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(471..487)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(480..502)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      629..1084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(716..735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     926..947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1110..1214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1215..1283,1284..2075)
                     /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      2246..2834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2735..2754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    3122..3143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3158..3163,3164..3181,3182..3188)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3196..3212
                     /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     3201..3223
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3220..3236
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3220..3236
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   3274..3403
                     /label=AAV2 ITR
     intron          4254..4386
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4369..4388
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4431..4449
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6347..6373
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6377..6433
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6443..7153
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6588..6606)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6856..6875
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 tctagagtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
       61 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
      121 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
      181 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
      241 acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat
      301 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg
      361 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
      421 cgggcggcct cagtgagcga gcgagcgcgc agccttaatt aacctaattc actggccgtc
      481 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      541 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      601 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
      661 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
      721 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
      781 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
      841 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
      901 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
      961 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1021 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1081 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1141 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1201 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1261 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1321 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1381 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1441 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1501 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1561 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1621 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1681 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1741 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1801 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1861 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1921 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1981 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2041 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2101 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2161 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2221 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2281 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2341 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2401 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2461 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2521 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2581 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2641 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2701 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2761 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2821 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2881 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2941 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3001 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3061 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3121 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     3181 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     3241 attacgccag atttaattaa ggccttaatt aggctgcgcg ctcgctcgct cactgaggcc
     3301 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     3361 gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac
     3421 ccgccatgct acttatctac gtagccatgc tctaggaaga tctaacatat cctggtgtgg
     3481 agtaggggac gctgctctga cagaggctcg ggggcctgag ctggctctgt gagctgggga
     3541 ggaggcagac agccaggcct tgtctgcaag cagacctggc agcattgggc tggccgcccc
     3601 ccagggcctc ctcttcatgc ccagtgaatg actcaccttg gcacagacac aatgttcggg
     3661 gtgggcacag tgcctgcttc ccgccgcacc ccagcccccc tcaaatgcct tccgagaagc
     3721 ccattgagca gggggcttgc attgcacccc agcctgacag cctggcatct tgggataaaa
     3781 gcagcacagc cccctagggg ctgcccttgc tgtgtggcgc caccggcggt ggagaacaag
     3841 gctctattca gcctgtgccc aggaaagggg atcaggggat gcccaggcat ggacagtggg
     3901 tggcaggggg ggagaggagg gctgtctgct tcccagaagt ccaaggacac aaatgggtga
     3961 ggggagagct ctccccatag ctgggctgcg gcccaacccc accccctcag gctatgccag
     4021 ggggtgttgc caggggcacc cgggcatcgc cagtctagcc cactccttca taaagccctc
     4081 gcatcccagg agcgagcaga gccagagcag gttggagagg agacgcatca cctccgctgc
     4141 tcgcaagctt tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgcttctg
     4201 acacaacagt ctcgaactta agctgcagaa gttggtcgtg aggcactggg caggtaagta
     4261 tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa
     4321 gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt gcctttctct
     4381 ccacaggtgt ccactcccag ttcaattaca gctcttaagg ctagagtact taatacgact
     4441 cactataggc tagcctcgag aattcatgac tgcggagcaa gcgctgcccc tgggcaacgg
     4501 gaaagcggcc gaggaggcgc gggggtccga gacgctgggc ggcggcggcg ggggcgcggc
     4561 ggggacgcgc gaggcgcgcg acaaggctgt ccacgagcgc ggccactgga acaacaaggt
     4621 ggagttcgtg ttgagcgtag ctggagagat catcggactg ggcaacgtgt ggcgctttcc
     4681 ctacctgtgc tacaaaaacg gcggaggggc attcctgatt ccttacgtgg tgtttttcat
     4741 ctgctgcgga atccccgtct tcttcctgga aacggctctg gggcagttca cgagcgaggg
     4801 tggcatcacg tgctggagga gggtctgccc tttatttgaa ggcattggct atgcaacaca
     4861 ggtgatcgag gcacacctca atgtctacta catcatcatc ctggcgtggg ccatcttcta
     4921 cctaagcaat tgcttcacca ccgagctccc ctgggccacc tgtgggcacg agtggaacac
     4981 agagaaatgt gtggagttcc agaagctgaa cttcagcaac tacagtcatg tgtccctgca
     5041 gaatgccacc tccccggtca tggagttctg ggaacgccgg gtcttggcta tatccgatgg
     5101 cattgagcac attgggaacc tccgttggga actggcatta tgtctcctgg cggcctggac
     5161 catctgctac ttctgcatat ggaagggtac taagtcgact ggaaaggtcg tgtatgtcac
     5221 tgcaaccttc ccctacatca tgctgctgat cctcctgatc cgaggggtca cgttgccagg
     5281 cgcctctgaa ggcatcaagt tctatctgta ccctgacctc tcccggctct ctgacccaca
     5341 ggtgtgggtg gacgctggga cgcagatctt tttctcctat gccatctgcc tgggctgcct
     5401 aaccgctctg gggagttaca acaactataa caacaactgc tacagggact gcattatgct
     5461 ctgctgtcta aacagtggca ccagcttcgt ggctgggttt gccatcttct cagtcctggg
     5521 cttcatggca tacgagcagg gtgtgcctat tgccgaggtg gcagagtcag gtcctggact
     5581 ggctttcatc gcctacccca aggctgtcac tatgatgccc ctctctccac tgtgggccac
     5641 cttgttcttc atgatgctca tcttcctggg cctggacagt cagtttgtgt gcgtggagag
     5701 tcttgtgaca gccgtggtag acatgtaccc taaggtcttc cggcggggct accggcgaga
     5761 actgctcatc ctggccctgt ccatcatctc ctatttccta ggcctggtga tgctgaccga
     5821 gggaggcatg tacatcttcc agctttttga ctcatacgcc gccagtggca tgtgcttgct
     5881 ctttgtggcc atctttgagt gtgtctgcat cggctgggta tatggaagca accggttcta
     5941 tgacaatatt gaggatatga ttggttaccg gccactgtca ctcatcaagt ggtgctggaa
     6001 agtcgtgacc cctgggatct gtgcgggcat cttcatcttc tttctggtca agtacaaacc
     6061 gctcaagtac aacaatgtgt acacatatcc tgcttggggc tacggcatcg gctggctcat
     6121 ggctctgtcc tccatgctgt gtatcccact ctggatcttc attaagctgt ggaagacaga
     6181 gggcacactg cccgagaaat tacagaagtt gacagtcccc agcgccgatc tgaaaatgcg
     6241 gggcaagctt ggggccagcc cacggacggt gaccgttaat gactgtgagg ccaaggtcaa
     6301 gggcgacggc accatctctg ccatcacaga gaaggagaca cacttctacc catacgatgt
     6361 tccagattac gctggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga
     6421 gaatcctgga ccgaccgagt acatggtgag caagggcgag gaggataaca tggccatcat
     6481 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     6541 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     6601 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     6661 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     6721 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     6781 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     6841 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     6901 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     6961 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     7021 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     7081 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     7141 gctgtacaag taa
//
