LOCUS       sequence-49611-e        7222 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 7222)
  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 7222)
  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-VMAT2-WPRE-pA
FEATURES             Location/Qualifiers
     source          1..7222
                     /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)."
     CDS             complement(2855..2884)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3175..3184
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3207..3240
                     /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    3291..3324
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3349..3937
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3402..3422)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3820..3831)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3969..4445
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4168..4187)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4485..4625
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4485..4614
                     /label=AAV2 ITR
     rep_origin      4700..5155
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4787..4806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4997..5018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5172..5191)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5291..5313
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5351..5369)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5437..5541
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5542..5610,5611..6402)
                     /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(5760..5779)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6573..7161
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7062..7081
                     /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 tagatttagc cacctcagtc actctcagat tcttcatcat
     1621 caccgacggg gtacggctgg acattattct gagtgtacat ctttgtctta atggggcagt
     1681 tgtggtccat gaggatagcc attttttctt ccttagcagg tggacttcga agaaaaaagc
     1741 agagtggagc aaaaacaata tcaattatcc caataattgt catgagccaa ggaaagccaa
     1801 ttgcctttgc aatagcacca ccagcagagg gaccgatagc ataccccatg cagaaggcta
     1861 catctgcaat ggcatacaca ctcccataga cagacacgtg ccgcaggtcc accaggtagc
     1921 ccatgatagg catcatggag gagtccacca tcccaatggc aaaaccaact ccaaagttgg
     1981 gagcgatgag tccgtagata ttttttgcaa aaggaataca taaaatgctg atcccaacaa
     2041 ctatcattcc cagaagagca cacagccacc ttcccatttt gtgtgcaagt atcccaaaaa
     2101 tattggttcc aatgagataa gagatgctcg ctggcaggaa ggcaacgccc agctgccact
     2161 ttcgggaaca catggtctcc atcatccaga tgggcagggc gggctccagc atggctatcc
     2221 ccatgtttgc aaagcagatg gagcctgcag cgatgagaat gtatgggtcc ttcagcaagg
     2281 tcgtaagagg tgtccccttc tgactttctg gctgcactcg ggatggctgg agcacaaaga
     2341 gctgaatagc tccatccaag agcaccaagg cagcaagcac caggaaagga gctgtcttcc
     2401 ccacaaactc atagagcaca cttccaaagg ggggtcccac taagaccccc atggccaggc
     2461 cccccagagc aatccccatg gcgttacccc tctcttcatc atctgtgtac acgctggcca
     2521 gcatgcccat cccagccaca gatgagcaag aggagccgat tccctgcagg gacctggcga
     2581 tcagcaggaa ggcatagctg ctggagaagg caaacataac tgttgagata aacataatgc
     2641 agaatccagc aaacatggga attggatagc caattctgtt ggttagaagt cctatgaatg
     2701 ggttggtgag gagctggaca gtggctttgg aggcaaacag cagcccaact tgcacattct
     2761 catttagaag gtctttgtct tcactggggc agtcgggagg gacagtcgtg ttggtcacag
     2821 tgtgctgtgt ggtggtagcc ttgggtgact cccccagatc ttcttcagaa ataagttttt
     2881 gttctcctgg gaggcccccc gtggcattcc ccgtgaacac agtagagttg ttatagtaag
     2941 agaagatgct gtggaagctt tcggaagtgg aggctgtgag cgctggcctg gcagtctgga
     3001 tttccgtagt atttttctcg tgcttaatgc tgtacaggta gctgggaatg atgggaacta
     3061 cgacggtgag cagcatgttg tctagcagca gcgcgaggaa cacgatgaac aggatcagct
     3121 tgcgcgagtg gcggctgtcc cgcagccatc gcagcagcac cagatcgctc agggccatgg
     3181 tggcggtggc gaccggtagc gctagcataa cttcgtataa agtatcctat acgaagttat
     3241 ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga ataacttcgt
     3301 ataatgtatg ctatacgaag ttatgaattc gatatcaagc ttatcgataa tcaacctctg
     3361 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3421 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3481 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3541 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3601 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3661 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3721 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     3781 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3841 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3901 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     3961 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     4021 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     4081 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     4141 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     4201 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     4261 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     4321 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     4381 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     4441 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     4501 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4561 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     4621 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4681 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4741 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4801 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4861 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4921 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4981 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5041 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5101 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     5161 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5221 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5281 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5341 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5401 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5461 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5521 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5581 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5641 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5701 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5761 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5821 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5881 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5941 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6001 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6061 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6121 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6181 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6241 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6301 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6361 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6421 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6481 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6541 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6601 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6661 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     6721 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6781 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6841 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6901 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6961 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7021 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7081 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7141 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7201 tgctggcctt ttgctcacat gt
//
