LOCUS       sequence-379297-        6386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-vector for optogenetic long-term silencing of GABAergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  AUTHORS   Wiegert Lab
  TITLE     Wiegert Lab Plasmids 2022
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6386)
  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: 
            "Unpublished"
COMMENT     Sequence Label: mDlx-Aion-citrine
FEATURES             Location/Qualifiers
     source          1..6386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          839..971
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     954..973
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1013..1022
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1958..1960,1961..1963,1964..2674)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2003..2024)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2264..2283)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2611..2632
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2722..3310
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2775..2795)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3193..3204)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3479..3560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3516..3535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3570..3589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3639..3779
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3639..3768
                     /label=AAV2 ITR
     rep_origin      3854..4309
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3941..3960)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4151..4172
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4326..4345)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4445..4467
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4505..4523)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4591..4695
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4696..4764,4765..5556)
                     /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(4914..4933)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5727..6315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6216..6235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6377..6386,1..120)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
       61 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      121 gcggccgcac gcgtttaatt aagaattcta tacactcaca gtggtttggc atatatttgg
      181 tgaaattttt taaggaaaaa ttagtgttgg tttcgatata tggtagcttt ttctctaaca
      241 taatttgaat aattcagcaa agccctacta ccagctgtac ttctgcagcc tcttccattc
      301 tttccagcat tataattttg gttaattttc aattttaggt cctacgtctc tgcaatttgt
      361 gtatgaataa cagaataatt tccctctttt gtttcgcctt tcctgttcct gaatctaaat
      421 aaagatggct ttttagtatt aaaagtggaa gaaaattaca ggtaattatc tttgacggta
      481 aaaacgctgt aatcagcggg ctacatgaaa aattactcta attatggctg catttaagag
      541 aatggaaaaa aaccttcttg tggataaaaa ccttaaattg tccccaatgt ctgcttcaaa
      601 ttggatggca ctgcagctgg aggctttgtt cagaattgat cctggggagc tacgaaccca
      661 aagtttcaca gtaggaagct tgatatcgaa ttcctgcagc ccgggctggg cataaaagtc
      721 agggcagagc catctattgc ttacatttgc ttctagcctg caggtcgagg agcgcagcct
      781 tccagaagca gagcgcggcg ccttaagctg cagaagttgg tcgtgaggca ctgggcaggt
      841 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
      901 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
      961 tctctccaca ggtgtccact cccagttcaa ttacagctct taagaaacta gcgccaccat
     1021 ggactatggc ggcgctttgt ctgccgtcgg actcttccaa acctcataca ctcttgagaa
     1081 caatggttct gttatttgca tcccgaacaa cggccagtgc ttctgcttgg cttggcttaa
     1141 atccaacgga acaaatgccg agaagttggc tgccaacatt ctgcagtgga tttcttttgc
     1201 gctttcagcg ctctgcctga tgttctacgg ctaccagacc tggaagtcta cttgcggctg
     1261 ggagaacatt tacgtggcca cgatccagat gatcaagttc atcatcgagt atttccatag
     1321 ctttgacgaa cctgcggtga tctactcatc caacggcaac aagacccggt ggcttcgtta
     1381 cgcgagctgg ctgctgaccg cccctgtcat tcttatccac ctgagcaacc ttacgggtct
     1441 ggcgaacgac tataacaagc gtaccatggg tctgctggtg tcatgcatcg gcggcatcgt
     1501 gtgggccacc acggccgcgc tgtccaaggg atacgtccgt gtcattttct tcctgatggg
     1561 cctgtgctac ggcatctaca cattcttcaa cgcagccaag gtctacattg aggcgtacca
     1621 caccgtgccg aagggccggt gtcgccaggt ggtgactggc atggcttggc tcttcttcgt
     1681 atcatggggt atgttcccca tcctgttcat cctcggcccc gagggcttcg gcgtcctgag
     1741 ccggtacggc tccaacgtcg gccacaccat cattgacctg atgtcgaagc agtgctgggg
     1801 tctgctcggc cactacctgc gcgtgctgat ccacagccat atcctcatcc acggcgacat
     1861 tcgcaagacc accaaattga acattggtgg cactgagatt gaggtcgaga cgctggtgga
     1921 ggacgaggcc gaggctggcg cggtcgcggc actcgagatg gtgagcaagg gcgaggagct
     1981 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2041 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2101 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
     2161 cctgatgtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2221 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2281 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2341 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2401 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2461 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2521 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     2581 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2641 cgggatcact ctcggcatgg acgagctgta caagtaaagg cgcgccaccc ctgcagggaa
     2701 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     2761 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2821 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2881 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2941 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3001 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3061 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3121 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     3181 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3241 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3301 gcctccccgc atcgataccg agcgctgctc gagagatctg tgatagcggc catcaagctg
     3361 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3421 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3481 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3541 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatcagc
     3601 ttatcgatac cgcatgcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     3661 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3721 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     3781 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     3841 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3901 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     3961 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4021 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4081 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4141 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4201 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     4261 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     4321 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4381 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4441 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4501 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4561 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4621 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4681 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4741 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4801 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4861 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4921 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4981 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5041 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5101 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5161 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5221 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5281 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5341 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5401 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5461 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5521 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5581 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5641 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5701 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5761 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5821 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5881 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5941 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6001 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6061 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6121 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6181 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6241 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6301 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6361 ccttttgctc acatgtcctg caggca
//
