LOCUS       sequence-451254-        6348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with Ple389 (ADORA2A MiniPromoter) driving expression of
            EmGFP. Contains WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6348)
  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 6348)
  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: pEMS2387
FEATURES             Location/Qualifiers
     source          1..6348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR
     intron          1598..1730
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1713..1732
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1751..1760
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1757..1759,1760..1762,1763..2476)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1802..1823)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2063..2082)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2410..2431
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2542..2562)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2960..2971)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3092..3213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3103..3122)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3157..3176
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3409..3538
                     /label=AAV2 ITR
     primer_bind     complement(3557..3574)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3557..3573)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3566..3588)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3715..4170
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3802..3821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4012..4033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4196..4300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4301..4369,4370..5161)
                     /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(4519..4538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5332..5920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5821..5840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6208..6229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6244..6249,6250..6267,6268..6274)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6282..6298
                     /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     6287..6309
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6306..6322
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6306..6322
                     /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 aggccggcca ggaggggctg agtaaatctg
      241 cgcctgattt tcatctgcct acactgaatt atgccatcca tttggaggca attttcaagt
      301 taaaatgttg tcagaaggct gtcacagttt ttatctttct tttctgttct ttctcaaccc
      361 acccacattt tctagttagg ttggaaaggg gagtgagggg gatagagcct ggtttatatt
      421 cagagatttc ttaacagggc cagaacccag gaagggatat gcttttctca ggaataatgt
      481 caaaattaat atagcttccc agtcaatgag gactatataa attactattt gcaaagcact
      541 aagaacagtg tccggcacag agtcagtgct atgtaatgta gatgtgagaa tgagagagcc
      601 tgagatttta tgtttgcgag tcccacagga ggcagggcaa tgccaactcc ctgttgcttg
      661 gaagcaatgg gcaagtctat gaccctcctg ttgaggccct gttacaggta accaagcccc
      721 cacggaatgg agctgaggag ctcctgagaa tgggcttgtc cggttactgt ttggattcag
      781 aactgaccgt ccacggccac ggtgtcagtc cctccttgtt ctgcgcttgc tttttggggt
      841 ctcccagcct ctccctagtc ctagagtttc tcaggcaaat cccagccact acccctccat
      901 ccgtgtcccc tgtttcccca gtgctgagct ggctgagctg tgagctttcc tgctccttgc
      961 ctcaggaagc caggggcagc ccagtcacct ctggccagag gcgggcctta ggccaagcct
     1021 ggttccgtac ctgctttctg ccagggacca aactcccgtt agtcattcct ggtgtcagcc
     1081 aggcagagga gcaggtgggg gagagggcgt gggaggaggg aggtgaggca gctccctccg
     1141 agtggaagct cacagtcagc tgcagcgccg cccctgccct gactgcagct gactggcagg
     1201 ggcacctgga ggcagcgggt tcgggcgcag tatgagaggg cttaccctct gcggcagtgg
     1261 gagacaggac caggcagctc cccacatcct tccacatccg agctccaggg gtctctggct
     1321 tgtcctttca cagggcggct ggtgtccctt aggtgggtgc tggtagttcc tgggagagtt
     1381 gacgggatgg agcaggccag gggggctgag aagttggcct ggcgagagaa ggtgggtggg
     1441 actgggagag cctgggcagg gaggggcctt tctgggaggg agtgtcatgg ggctggccga
     1501 gatccaggac ccaggagact ggccaagctg ccacaggagg cctgagaggc tgagcatggg
     1561 gggggcgcgc cgaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
     1621 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
     1681 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
     1741 ccaggcggcc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1801 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1861 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1921 cgtgccctgg cccaccctcg tgaccacctt cacctacggc gtgcagtgct tcgcccgcta
     1981 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2041 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2101 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2161 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaaggtct atatcaccgc
     2221 cgacaagcag aagaacggca tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg
     2281 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2341 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     2401 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2461 cgagctgtac aagtaagcgg ccgcgatcgc tcaacctctg gattacaaaa tttgtgaaag
     2521 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2581 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2641 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2701 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2761 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2821 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2881 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2941 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3001 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3061 ttgggccgcc tccccgcgat cgcagacatg ataagataca ttgatgagtt tggacaaacc
     3121 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3181 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3241 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3301 ggtagtcgaa attcccgata aggatcttcg tcgacgaatt cccgataagg atcttcctag
     3361 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     3421 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3481 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc
     3541 ttaattaacc taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     3601 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3661 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     3721 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3781 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3841 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3901 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3961 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4021 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4081 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4141 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     4201 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4261 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4321 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4381 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4441 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4501 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4561 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4621 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4681 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4741 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4801 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4861 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4921 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4981 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5041 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5101 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5161 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5221 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5281 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5341 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5401 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5461 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5521 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5581 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5641 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5701 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5761 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5821 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5881 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5941 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6001 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6061 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6121 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6181 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6241 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6301 tcacacagga aacagctatg accatgatta cgccagattt aattaagg
//
