LOCUS       sequence-49597-e        7204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7204)
  AUTHORS   Tritsch NX, Ding JB, Sabatini BL
  TITLE     Dopaminergic neurons inhibit striatal output through non-canonical 
            release of GABA.
  JOURNAL   Nature. 2012 Oct 11;490(7419):262-6. doi: 10.1038/nature11466. Epub 
            2012 Oct 3.
  PUBMED    23034651
REFERENCE   2  (bases 1 to 7204)
  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: 
            "10.1038/nature11466"; journalName: "Nature"; date: "2012-10-11- 
            11"; volume: "490"; issue: "7419"; pages: "262-6"
COMMENT     Sequence Label: pAAV-Ef1a-DIO-VGAT-WPRE-pA
FEATURES             Location/Qualifiers
     source          1..7204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1450..1483)
                     /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(1534..1567)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3163..3172
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3189..3222
                     /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    3273..3306
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3331..3919
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3384..3404)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3802..3813)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3951..4427
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4150..4169)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4467..4607
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4467..4596
                     /label=AAV2 ITR
     rep_origin      4682..5137
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4769..4788)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4979..5000
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5154..5173)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5273..5295
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5333..5351)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5419..5523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5524..5592,5593..6384)
                     /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(5742..5761)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6555..7143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7044..7063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1561 aagttatggc gcgccttatc tagatttact agtcctctgc gttggttcgg taggcctcga
     1621 tgagaccctc gagtgaatgc acgaagccgg acacgctgca gatgccgccg atgacgaaga
     1681 tggccacatc gaagaagacc tggtgccaca gcagcttgcg ccagagaagg cgcaagtgga
     1741 agaggctggg cagcaggaag cagaggccgg cgcctgtgag gctgcccgtg aggcccatga
     1801 gcagcgcgaa gtgtggcacg tagatggcca tgagcagcgt gaagaccacc agcgcgcagc
     1861 gcagcgtcag cccccaggac ttaaggcgac cgtcgcctcc atagcaggcg gggaagaagg
     1921 cgcgactgcc ttcctggaag agagacttct ccagcacttc gacggccgcg aagaagggca
     1981 acggatagga cagcagcgcc ttggccacca ggaagaggtt gaccacggcg cggatggagc
     2041 cgggcaggtt atccgtgatg acttccttgg tctcgtcggc ccaggtgagg taggcgacga
     2101 gcgcgaagag acccttgagc acgcaggcgg cgatgtgtgt ccagttcatc atgcagtgga
     2161 attcgctggg ctgctgcatg ttgccttcga gagagggcag gaagatctgc gacgtgtagc
     2221 tgaacacgat gatgccaatg gagatgggaa acttcttgac gtcgatgtag aacttcacct
     2281 tctcccaggc ccaatcacgc gcgcgagaga gacagtaagc gatgaccagg atgttgatga
     2341 cgaagtgggc cagcgtacac agcagactga acttggacac ggccttgaga ttcttcagga
     2401 aggcgcaggg cagcagcacc gctgtggcta tgatggacca ggacttctgc gacacgggca
     2461 gccccgggaa actgttgtac atgaggttgc cgctcaccac gacgtacaag atacacgtca
     2521 tcaccagctc gatgatctgc gccacattga ccacgcggcc gcccagggtg gggaatcgag
     2581 gagcgcagca tgcgttagct atggccacat acgagtcccg cacgcgcacc acctccccgt
     2641 cttcgttctc ctcgtacagg cacgcgatga ggatcttgcc ggtgtagcag cacaccactg
     2701 cggcgaagat gatgaggaac aaccccaggt agccgccgtg gaggatggcg tagggtaggc
     2761 ccagcacgaa catgccctga atggcatttg tcacgttcca gcccgcttcc cacgccgtga
     2821 tcttgggttt gtcgtgaccc ccgaactccc caccagctcc cacggcctgg tccttggagc
     2881 cagagggtgg cagaggagcg ccgccgcgct gataatgaat gtctccctcg acgggagctt
     2941 ctgcgccctc gtctccgcag ggctcgcctt ccgatttcag gatgtccatc tgcaggccct
     3001 ggcgatgctc aaagtcgaga tcgtcgcagt gcgcgaagcc caccgcttcc tcatcggtgg
     3061 ccgcctgaaa ccccatcctg gcaaacatgc cgctcacctt ggcctgggac ttgttggaca
     3121 cggaggtggc cacattggtc agcttgctgc ggagcagggt ggccatggtg gcgaccggta
     3181 gcgctagcat aacttcgtat aaagtatcct atacgaagtt atttgcctta acccagaaat
     3241 tatcactgtt attctttaga atggtgcaaa gaataacttc gtataatgta tgctatacga
     3301 agttatgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     3361 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3421 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3481 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3541 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3601 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3661 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3721 gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg
     3781 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3841 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3901 ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc
     3961 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     4021 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     4081 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     4141 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     4201 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     4261 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     4321 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     4381 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg
     4441 taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     4501 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4561 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     4621 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     4681 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4741 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4801 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4861 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     4921 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4981 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
     5041 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5101 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     5161 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     5221 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5281 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5341 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5401 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5461 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5521 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5581 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5641 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5701 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5761 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5821 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5881 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5941 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6001 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6061 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6121 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6181 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6241 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6301 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6361 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6421 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6481 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6541 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6601 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6661 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     6721 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6781 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6841 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6901 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6961 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7021 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7081 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7141 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7201 atgt
//
