LOCUS       sequence-403390-        6150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  non-functional Kir2.1(mutant) expression as a control for the 
            silencing of neurons with Kir2.1 and mScarlet-KASH expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6150)
  AUTHORS   Roethler O, Zohar E, Cohen-Kashi Malina K, Bitan L, Gabel HW, 
            Spiegel I
  TITLE     Single genomic enhancers drive experience-dependent GABAergic 
            plasticity to maintain sensory processing in the adult cortex.
  JOURNAL   Neuron. 2023 Jun 20:S0896-6273(23)00399-9. doi: 
            10.1016/j.neuron.2023.05.026.
  PUBMED    37354902
REFERENCE   2  (bases 1 to 6150)
  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. 
            2023 Jun 20:S0896-6273(23)00399-9. doi: 
            10.1016/j.neuron.2023.05.026."
COMMENT     Sequence Label: pAAV-hSyn-Kir2.1Mut-2A-mScarlet-KASH
FEATURES             Location/Qualifiers
     source          1..6150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        162..609
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1934..1987
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      1997..2006
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2003..2698
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2136..2154)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2404..2423
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3038..3058)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3418..3558
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3418..3547
                     /label=AAV2 ITR
     rep_origin      3633..4088
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3720..3739)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3930..3951
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4105..4124)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4224..4246
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4284..4302)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4370..4474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4475..4543,4544..5335)
                     /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(4693..4712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5506..6094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5995..6014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcagatgct gcatgcgtat gagtgcaagt gggttttagg
      181 accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag
      241 cacccaaccc ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg
      301 atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg
      361 cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt
      421 cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc
      481 ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc
      541 gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg
      601 agagcgcagg gtaccggatc ctctagagtc gactgccacc atgggcagtg tgagaaccaa
      661 ccggtacagc attgtctctt cggaggaaga tggcatgaag ctggccacta tggcagttgc
      721 caatggcttt gggaatggca agagtaaagt ccatacccga caacagtgca ggagccgctt
      781 tgtgaagaaa gatggtcatt gcaatgttca gtttatcaac gtgggtgaga agggacagag
      841 gtacctggca gacatcttta ctacctgtgt cgacatccgc tggaggtgga tgctggttat
      901 cttctgtctt gccttcgtgc tctcctggct gttctttggc tgtgtgtttt ggttgatagc
      961 cctgctccat ggggatctag atacttctaa agtgagcaaa gcatgcgtgt cggaggttaa
     1021 cagcttcacg gctgccttcc tcttctccat cgagacccag acaaccattg ccgctgcttt
     1081 caggtgtgtg acagacgaat gcccaattgc tgtcttcatg gtggtattcc agtcaattgt
     1141 aggctgcatc attgacgcct tcatcattgg tgcagtcatg gcgaagatgg caaagccaaa
     1201 gaagagaaat gagactcttg tcttcagtca caatgctgtg attgccatga gggatggcaa
     1261 actctgcttg atgtggagag tgggtaacct tcgaaagagc caccttgtgg gagctcatgt
     1321 ccgggcacag cttctcaaat ctaggatcac ttcagaaggg gagtttatcc ctttggacca
     1381 gatagacatc aatgttggtt ttgatagtgg aattgaccgc atatttctag tgtcccccat
     1441 cactatcgtt cacgaaatag atgaagacag ccctttatat gacttgagta agcaggacat
     1501 tgacaatgca gactttgaaa ttgttgtcat actggaaggc atggtggagg cgactgccat
     1561 gacaactcaa tgccggagtt cgtatctggc caatgaaatt ctctggggtc accgctatga
     1621 gccagtgctc tttgaagaga aacactacta taaagtagac tattcaagat tccataagac
     1681 ttatgaagta cctaacaccc ccctttgtag tgccagagac ttagcagaga agaaatacat
     1741 cctttcaaat gcaaattcat tttgctatga aaatgaagtt gccctaacaa gcaaagagga
     1801 agaggaggat agtgagaacg gagtcccaga gagcacaagc acagactcac ctcctggcat
     1861 agatctccac aaccaggcaa gcgtacctct agagcccagg cccttaaggc gagaatcgga
     1921 gataggcagt ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc
     1981 tggcccagcg gatcccgcca ccatggtgag caagggcgag gcagtgatca aggagttcat
     2041 gcggttcaag gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga
     2101 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     2161 ccccctgccc ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt
     2221 caccaagcac cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa
     2281 gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc
     2341 cctggaggac ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga
     2401 cggccccgta atgcagaaga agacaatggg ctgggaagcg tccaccgagc ggttgtaccc
     2461 cgaggacggc gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggacg
     2521 ctacctggcg gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc
     2581 ctacaacgtc gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga
     2641 acagtacgaa cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc
     2701 cggactcaga tctcgagagg aggaggagga gacagacagc aggatgcccc acctcgacag
     2761 ccccggcagc tcccagccga gacgctcctt cctctcaagg gtgatcaggg cagcgctacc
     2821 gttgcagctg cttctgctgc tgctgctgct cctggcctgc ctgctacctg cctctgaaga
     2881 tgactacagc tgcacccagg ccaacaactt tgcccgatcc ttctacccca tgctgcggta
     2941 caccaacggg ccacctccca cctaagaatt cgatatcaag cgataatcaa cctctggatt
     3001 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3061 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3121 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     3181 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg tttatttgtg
     3241 aaatttgtga tgctattgct ttatttgtaa ccatctagct ttatttgtga aatttgtgat
     3301 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3361 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag cggccgcagg
     3421 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3481 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3541 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3601 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3661 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     3721 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3781 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3841 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3901 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3961 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     4021 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     4081 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4141 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4201 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4261 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4321 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4381 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4441 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4501 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4561 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4621 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4681 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4741 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4801 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4861 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4921 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4981 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5041 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5101 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5161 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5221 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5281 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5341 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5401 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5461 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5521 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5581 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5641 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5701 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5761 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5821 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5881 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5941 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6001 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6061 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6121 gttcctggcc ttttgctggc cttttgctca
//
