LOCUS       sequence-429066-        6150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kir2.1 expression for silencing neurons 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-Kir2.1WT-2A-mScarlet-KASH
FEATURES             Location/Qualifiers
     source          1..6150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   13..142
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        169..616
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      642..651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1941..1994
                     /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      2004..2013
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2010..2705
                     /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(2143..2161)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2411..2430
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3045..3065)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3425..3565
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3425..3554
                     /label=AAV2 ITR
     rep_origin      3640..4095
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3727..3746)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3937..3958
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4112..4131)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4231..4253
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4291..4309)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4377..4481
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4482..4550,4551..5342)
                     /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(4700..4719)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5513..6101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6002..6021
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
       61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctgcggccgc agatgctgca tgcgtatgag tgcaagtggg
      181 ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact
      241 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg
      301 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc
      361 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact
      421 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg
      481 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg
      541 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
      601 gtgcctgaga gcgcagggta ccggatcctc tagagtcgac tgccaccatg ggcagtgtga
      661 gaaccaaccg gtacagcatt gtctcttcgg aggaagatgg catgaagctg gccactatgg
      721 cagttgccaa tggctttggg aatggcaaga gtaaagtcca tacccgacaa cagtgcagga
      781 gccgctttgt gaagaaagat ggtcattgca atgttcagtt tatcaacgtg ggtgagaagg
      841 gacagaggta cctggcagac atctttacta cctgtgtcga catccgctgg aggtggatgc
      901 tggttatctt ctgtcttgcc ttcgtgctct cctggctgtt ctttggctgt gtgttttggt
      961 tgatagccct gctccatggg gatctagata cttctaaagt gagcaaagca tgcgtgtcgg
     1021 aggttaacag cttcacggct gccttcctct tctccatcga gacccagaca accattggct
     1081 atggtttcag gtgtgtgaca gacgaatgcc caattgctgt cttcatggtg gtattccagt
     1141 caattgtagg ctgcatcatt gacgccttca tcattggtgc agtcatggcg aagatggcaa
     1201 agccaaagaa gagaaatgag actcttgtct tcagtcacaa tgctgtgatt gccatgaggg
     1261 atggcaaact ctgcttgatg tggagagtgg gtaaccttcg aaagagccac cttgtgggag
     1321 ctcatgtccg ggcacagctt ctcaaatcta ggatcacttc agaaggggag tttatccctt
     1381 tggaccagat agacatcaat gttggttttg atagtggaat tgaccgcata tttctagtgt
     1441 cccccatcac tatcgttcac gaaatagatg aagacagccc tttatatgac ttgagtaagc
     1501 aggacattga caatgcagac tttgaaattg ttgtcatact ggaaggcatg gtggaggcga
     1561 ctgccatgac aactcaatgc cggagttcgt atctggccaa tgaaattctc tggggtcacc
     1621 gctatgagcc agtgctcttt gaagagaaac actactataa agtagactat tcaagattcc
     1681 ataagactta tgaagtacct aacacccccc tttgtagtgc cagagactta gcagagaaga
     1741 aatacatcct ttcaaatgca aattcatttt gctatgaaaa tgaagttgcc ctaacaagca
     1801 aagaggaaga ggaggatagt gagaacggag tcccagagag cacaagcaca gactcacctc
     1861 ctggcataga tctccacaac caggcaagcg tacctctaga gcccaggccc ttaaggcgag
     1921 aatcggagat aggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     1981 agaatcctgg cccagcggat cccgccacca tggtgagcaa gggcgaggca gtgatcaagg
     2041 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
     2101 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     2161 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2221 gggccttcac caagcacccc gccgacatcc ccgactacta taagcagtcc ttccccgagg
     2281 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc gtgacccagg
     2341 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
     2401 ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc accgagcggt
     2461 tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc ctgaaggacg
     2521 gcggacgcta cctggcggac ttcaagacca cctacaaggc caagaagccc gtgcagatgc
     2581 ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag gactacaccg
     2641 tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg gacgagctgt
     2701 acaagtccgg actcagatct cgagaggagg aggaggagac agacagcagg atgccccacc
     2761 tcgacagccc cggcagctcc cagccgagac gctccttcct ctcaagggtg atcagggcag
     2821 cgctaccgtt gcagctgctt ctgctgctgc tgctgctcct ggcctgcctg ctacctgcct
     2881 ctgaagatga ctacagctgc acccaggcca acaactttgc ccgatccttc taccccatgc
     2941 tgcggtacac caacgggcca cctcccacct aagaattcga tatcaagcga taatcaacct
     3001 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3061 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3121 attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc
     3181 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttt
     3241 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca tctagcttta tttgtgaaat
     3301 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3361 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcgg
     3421 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3481 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3541 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3601 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3661 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3721 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3781 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3841 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3901 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3961 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4021 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4081 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4141 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4201 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4261 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4321 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4381 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4441 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4501 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4561 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4621 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4681 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4741 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4801 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4861 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4921 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4981 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5041 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5101 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5161 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5221 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5281 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5341 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5401 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5461 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5521 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5581 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5641 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5701 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5761 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5821 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5881 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5941 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6001 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6061 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6121 ttttacggtt cctggccttt tgctggcctt
//
