LOCUS       sequence-445483-        6366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with Ple385 (GPR6 MiniPromoter) driving expression of 
            EmGFP. Contains WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6366)
  AUTHORS   de Moura Gomes A, L Petkau T, J Korecki A, Fornes O, Galvan A, Lu G,
            M Hill A, Ling Lam S, Yao A, A Farkas R, W Wasserman W, Smith Y, M 
            Simpson E, R Leavitt B
  TITLE     New MiniPromoter Ple389 (ADORA2A) drives selective expression in 
            medium spiny neurons in mice and non-human primates.
  JOURNAL   Sci Rep. 2024 Nov 15;14(1):28194. doi: 10.1038/s41598-024-79004-y.
  PUBMED    39548191
REFERENCE   2  (bases 1 to 6366)
  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: "Sci Rep."; 
            date: "2024-11-15"; volume: "14(1):28194. doi"; pages: " 
            10.1038/s41598-024-79004-y"
COMMENT     Sequence Label: pEMS2383
FEATURES             Location/Qualifiers
     source          1..6366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR
     intron          1616..1748
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1731..1750
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1769..1778
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1775..1777,1778..1780,1781..2494)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1820..1841)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2081..2100)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2428..2449
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2560..2580)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2978..2989)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3110..3231
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3121..3140)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3175..3194
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3427..3556
                     /label=AAV2 ITR
     primer_bind     complement(3575..3592)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3575..3591)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3584..3606)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3733..4188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3820..3839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4030..4051
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4214..4318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4319..4387,4388..5179)
                     /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(4537..4556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5350..5938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5839..5858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6226..6247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6262..6267,6268..6285,6286..6292)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6300..6316
                     /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     6305..6327
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6324..6340
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6324..6340
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      181 agccatgctc taggaagatc ggaattccct aggccggccg aaaaatacct agtcttgaac
      241 tttggagttc caaattccac atgatcgact tcctcaaggc ttttcacttg acgaacaaat
      301 tgcatgccaa gaaatcgtca aaattctatc aagcttaatt atgcaatttc tgtctataaa
      361 tgacatacct cctgtaccct atgacaccca cctatgcact aataagatgt gacagctgca
      421 tcaccaacaa agcctccccc gcctgccatt ctcttcggag gtgagcacct cctgctcggg
      481 aggaactgac agagacagga ggaggtgggg acacagttcc cctactctga caggaggcaa
      541 ggtgggaata attggtactg atagatctaa gtttatggga ggtgggagga agttcataga
      601 gttcctacct gacaacctag attcatctga aggaggagtg ccagttaaga gactgttaca
      661 agaagtatcg agaacctgag ttgatgacat tgggaacaaa gctgtcccaa cctccccctt
      721 ctccggggtt tcttttttga cctgatccct cagtgatctt tgacgctggg ctttgctgcc
      781 cggcaggtgt cactcatcct tcacccctcc cctcccagct caccctcttc gccgcctggt
      841 ctcagggtct tcttcactcc tttctacctc tccttgtctc tctcccatcc cattcctgga
      901 agttctggac tggaggtggt tgctctggct tctcctccct cctaggaccc tcagatagtg
      961 ttgggggcag ggggagtgcg gcggggaggg cacaggatgg gctggcgagg agagtgagtg
     1021 ctgggctcgc accgggagcc gggagcctga cgtcaccacg ccctgcctgt caatctgcag
     1081 cgcgcgccgc tcgcagccgc cttttctgcc accaactgta tctctcactc gcggagccgg
     1141 cacagcgaca ggcgccccgc gcttgcctcc tccccggccc ggccgcctcc tgcaggcagc
     1201 tgctctgttt gcgcggggag ctcagcccgg cgccccgcct cggcgcccat gaccagcgac
     1261 ttccagaagt cctgacgccc caggtgagtg gccgagttcc cagggaactt tgaggatgtg
     1321 gggagaggag gggagaaaga tatccagggg acaggtcttt ccagaagagg agaaatggag
     1381 ccaaccttga ctcccaccct ctgctgcccc catacacaca ccctagtctc tcctcccaga
     1441 cccccttcgc aggggtctct gggctccagg actctccagg cttcgcaatg ccagaaactt
     1501 cgctttcagc accgcggaca gcgtctctgc tgcccagccc catggggatg tcctggtcgc
     1561 gccgccacaa cccaggcggg aggcgcgccg aagttggtcg tgaggcactg ggcaggtaag
     1621 tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag
     1681 aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
     1741 ctccacaggt gtccactccc aggcggccgc caccatggtg agcaagggcg aggagctgtt
     1801 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1861 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1921 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttca cctacggcgt
     1981 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2041 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2101 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2161 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2221 caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact tcaagacccg
     2281 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2341 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     2401 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2461 gatcactctc ggcatggacg agctgtacaa gtaagcggcc gcgatcgctc aacctctgga
     2521 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2581 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2641 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2701 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2761 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2821 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2881 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     2941 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3001 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3061 gacgagtcgg atctcccttt gggccgcctc cccgcgatcg cagacatgat aagatacatt
     3121 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3181 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3241 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     3301 taaaacctct acaaatgtgg tagtcgaaat tcccgataag gatcttcgtc gacgaattcc
     3361 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     3421 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3481 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3541 gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg
     3601 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     3661 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     3721 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3781 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3841 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3901 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3961 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4021 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4081 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4141 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     4201 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4261 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4321 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4381 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4441 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4501 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4561 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4621 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4681 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4741 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4801 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4861 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4921 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4981 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5041 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5101 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5161 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5221 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5281 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5341 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5401 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5461 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5521 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5581 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5641 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5701 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5761 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5821 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5881 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5941 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6001 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6061 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     6181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     6241 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     6301 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccagatttaa
     6361 ttaagg
//
