LOCUS       sequence-330486-        7671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expressing cre-dependent mGluR1-myc-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  AUTHORS   Cai Y, Nielsen BE, Boxer EE, Aoto J, Ford CP
  TITLE     Loss of nigral excitation of cholinergic interneurons contributes to
            parkinsonian motor impairments.
  JOURNAL   Neuron. 2021 Apr 7;109(7):1137-1149.e5. doi: 
            10.1016/j.neuron.2021.01.028. Epub 2021 Feb 17.
  PUBMED    33600762
REFERENCE   2  (bases 1 to 7671)
  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"; doi: 
            "10.1016/j.neuron.2021.01.028"; journalName: "Neuron"; date: 
            "2021-04-7- 7"; volume: "109"; issue: "7"; pages: "1137-1149.e5"
COMMENT     Sequence Label: AAV hSYN DIO mGluR1-myc-FLAG
FEATURES             Location/Qualifiers
     source          1..7671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    637..670
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    733..766
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(803..826)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(845..874)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3607..3616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3642..3674)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(3745..3778)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3798..4386
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3851..3871)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4269..4280)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4418..4894
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4617..4636)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4934..5074
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4934..5063
                     /label=AAV2 ITR
     rep_origin      5149..5604
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5236..5255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5446..5467
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5621..5640)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5740..5762
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5800..5818)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5886..5990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5991..6059,6060..6851)
                     /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(6209..6228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7022..7610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7511..7530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccgataa cttcgtataa agtatcctat
      661 acgaagttat atcaaaatag gaagaccaat gcttcaccat cgacccgaat tgccaagcat
      721 caccatcgac ccataacttc gtataatgta tgctatacga agttatgcta gcgtttaaac
      781 ttaagctata gaattcttaa accttatcgt cgtcatcctt gtaatccagg atatcatttg
      841 ctgccagatc ctcttctgag atgagtttct gctcgagcgg ccgcgtacgc gtaagctgca
      901 catgtgccga cggacactgg ctgctgggcg agaattctgg ctgcctcttc ttggcgttcc
      961 ctgccccagg cttctttctc cggaaaatgt tgaggaaggt gttggagcgg catggcagct
     1021 tgccgtcacc gacgtgcatg cgcactacat cagaggtggt gaaggcactg cggacattcc
     1081 tctcgggttt agcaataatg atgtacatct tgggagtgaa catgcagccc agggccaccg
     1141 tcacactgag gctcactgcg aagcaggtag tgataatctt gtagttgctc ccaaagtaaa
     1201 tgggaacaaa agccagccag atgatgcagg tggtgtacat agtgaaggcg atgtatttag
     1261 cctcattgaa attggccggc acgttgcggg tcttgaaggc atagtaggta cagctcatga
     1321 tgagaagtcc gttgtaaccc acaggtgcca ctacacccag gttgctggta ttgcagataa
     1381 gatagacttc cttgatgctc gggtaggaca aaatgggcat gggaggctcc atgatgatca
     1441 aggtcaccac tagtgtcagc tgtacactaa tcagaatgga ggctatgatc acttgggccc
     1501 aagcgctcat gaacctgggc ttccgggtac agatcttctt cttgctgcca gccaggatgc
     1561 gtgcaatacg attggttttg gtcacaagag cagagtagca catggcagaa gagaggccaa
     1621 ctaggaggcg ctggaggtag caggatgtgg tagtaggttt ggcgatgagg gtgaaagggc
     1681 atacgtagcc gaggaaaata ccagccagaa tgatatagca gagctctcta ctggaggatt
     1741 tgaccacagg tgtgtcccgg tacagcacaa agatgagggt gacaaatagc gtcacgagga
     1801 tgcccaggca agaaaaggcg atggctatga tggattctat gtcactccac tcgaggtaac
     1861 ggatagtaat gggctcacag cctgtgagct ctgcattggg ccaccacccc aggtcacagg
     1921 ctctgcaggt aaactcatct tgcacaaact cattctcttt gcaggctgtg cagatccagc
     1981 agcagctcac ttcccctttc cgtatgacct taatctgacc ctttaagcaa ggctcgctgc
     2041 acacagatcg taccattccg cttttgttca tctggatttt gtaatcatcg atattcagca
     2101 caccttcatg ccaggttccc acatggacat agtcatagcg attagcctct gtgtactgca
     2161 gattcataat gtcatacctt ccaggtgcat cccccttctc atcgaaccag acctcctctc
     2221 cagacactcc aacaaaagag gatttgatga ggaaatccag gagcttcctg ccgtcaatgg
     2281 gcttcatggc atcacaaagg cccacgtagc caggacatag agcatggtgc atgttctgta
     2341 ggccgtgtgc catggcatag atggcgttga tgacaaatcc cattttgctg tcctggacat
     2401 aattttcttc caagctttca tttcctgtgc agactttttt aaagttgggg ttttccaaga
     2461 ggtgtccagg taggcgacac tggaagcgat gttgccagaa ctcagggaac caaggattcc
     2521 tcgtgttggt gtccagcctc agcttcagaa agtagtcatc aaacgacctg acctctggag
     2581 actgcagctt tattgtgatt cccccattgg cttccacctc atagccttcg atgacttcat
     2641 ctctgtctgc ccatccatca cttccaatga gtgagaactc tcccacgacg ccaaggcggc
     2701 gcatggcact cagtaacccc cgcactgtca tgccctcgca gaagcagacc acaaccctgg
     2761 ccttgggaag ccgctctcgg agcttgcgca gaagccgatc aaagctcttc tcgccagcat
     2821 tgctgtagat tttgtccgag tgtgcgatgc agagtccttc ctgggcagcc aactctttga
     2881 aagcatccat tccactctcg ccgtaattcc cttctgtgtg gactgcagag acataggtcc
     2941 agttgtaccg cttgacgata tcaagcatcg cccttgcctg caaagtgtca gaagggacca
     3001 ccctcaggaa gtatttgtac aaagttttgt cgctcaggtc tatgctcgtg gcagaatagg
     3061 cgatttgtgg gatgtcgaac agctgcagaa gattctggac ttgaatggcc acagagctgg
     3121 agccagggcc gatcactcca gcaataggct tcttagtcct gcctgggggt agggtctggc
     3181 catcaggcag gcagcggttc agtccatcct tctcatctcg gatggaaatc agggagtctc
     3241 tgatgaactc aatgctctgt tccagagcca cagaagagtg ccagcaggag tcccggatct
     3301 cactgcccag agtgatgttg ggcaggagca ccgggtccgc attaatctta tctaatgtgt
     3361 ggaacatggc ctccactctc tggataccat actgttctct gatctcccca cacttccttt
     3421 ccggtacctt ttcggctgga ggctggtgat ggactgagaa gagggctccg atgatgacat
     3481 ctccgtccat tctcgccaca gagcgctggg acgaggcacc tgccagcagt acttttctgt
     3541 caggcatcct gggcaaaatg gacatctcca aaaagatcat tgggaagaaa atcaagagga
     3601 gccggaccat ggtggcggat ccctagaggg cccgtttaaa ctaacttcgt ataggatact
     3661 ttatacgaag ttatcattgg gattcttcct attttgatcc aagcatcacc atcgaccctc
     3721 tagtccagat ctcaccatcg acccataact tcgtatagca tacattatac gaagttatcg
     3781 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3841 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3901 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3961 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4021 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4081 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4141 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     4201 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     4261 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     4321 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     4381 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     4441 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     4501 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     4561 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     4621 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     4681 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     4741 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     4801 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     4861 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     4921 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4981 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     5041 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     5101 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     5161 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5221 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5281 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5341 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     5401 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5461 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     5521 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5581 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     5641 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5701 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5761 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     5821 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5881 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5941 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6001 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6061 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6121 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6181 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6241 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6301 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6361 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6421 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6481 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6541 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6601 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6661 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6721 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6781 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6841 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6901 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6961 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7141 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
