LOCUS       sequence-369061-        6905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Aldh1a1 fused to HA, driven by the Ef1a promoter, in a 
            Cre-dependent fashion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6905)
  AUTHORS   Melani R, Tritsch NX
  TITLE     Inhibitory co-transmission from midbrain dopamine neurons relies on 
            presynaptic GABA uptake.
  JOURNAL   Cell Rep. 2022 Apr 19;39(3):110716. doi: 
            10.1016/j.celrep.2022.110716.
  PUBMED    35443174
REFERENCE   2  (bases 1 to 6905)
  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: "Cell Rep.";
            date: "2022-04-19"; volume: "39(3):110716. doi"; pages: " 
            10.1016/j.celrep.2022.110716"
COMMENT     Sequence Label: pAAV-EF1a-double floxed-Aldh1a1-HA-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      87..542
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(174..193)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     384..405
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(559..578)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     678..700
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(738..756)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        824..928
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(929..997,998..1789)
                     /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(1147..1166)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1960..2548
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2449..2468
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2610..2739
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2840..4018
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3071..4009
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3966..3986
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(4049..4082)
                     /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(4133..4166)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4178..4267)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     protein_bind    5784..5817
                     /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    5868..5901
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5926..6514
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5979..5999)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6397..6408)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6548..6659
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   join(6777..6905,1..12)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(6777..6905,1)
                     /label=AAV2 ITR
ORIGIN
        1 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
       61 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
      121 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
      181 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
      241 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
      301 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
      361 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
      421 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
      481 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
      541 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
      601 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
      661 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
      721 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
      781 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      841 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      901 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      961 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1021 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1081 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1141 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     1201 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1261 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1321 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1381 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1441 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1501 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1561 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1621 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1681 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1741 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1801 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1861 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1921 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1981 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2041 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2101 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2161 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2221 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2281 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2341 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2401 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2461 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2521 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2581 ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac
     2641 tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
     2701 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg cgtaagcttt
     2761 gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt
     2821 cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     2881 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg
     2941 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     3001 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     3061 cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg
     3121 cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct
     3181 tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg
     3241 tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct
     3301 tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc
     3361 tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg
     3421 tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc
     3481 ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg
     3541 ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag
     3601 gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc
     3661 agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca
     3721 aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc
     3781 gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg
     3841 ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc
     3901 agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt
     3961 cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg
     4021 taccggatcc tctagagtcg actccggaat aacttcgtat aggatacttt atacgaagtt
     4081 atgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct taataacttc
     4141 gtatagcata cattatacga agttatggcg cgccttaagc gtaatctgga acgtcatatg
     4201 gataggatcc tgcatagtcc gggacgtcat agggatagcc agcgtaatct ggaacatcgt
     4261 atgggtagcc ggagttcttc tgagatatct tcattgcgac tgtcttgagc tcagtgtatt
     4321 cataaagacc atgttcaccc agttctcttc catttccaga catcttgaat ccaccgaagg
     4381 ggcactgggc tgacaacatc atatagcagt taacccacac caccccagcc tgcagagcag
     4441 atgacacagt gatggcctta tccaggtctt tagtgaagag tcctgctgct aaaccatagg
     4501 tagtattgtt tgctctcttg atcacatcat ctacagactt aaacttcatg atttgttgca
     4561 ctggtccaaa tatctcctct ttggcaatgc gcatctcatc agtcacgttg gagaacactg
     4621 tgggctgcac aaagaagcct ttgttccccc agcgtcctcc accacactcc agtttggctc
     4681 cttctttctt cccactctca atgagatcga gtattttatc atgttgttcc ttgtcaatct
     4741 gagggccttg atttattcct ggggtcagag gatttccaag aacatatttc ttagctcgct
     4801 caacactcct tttcacaaac tcatcataaa ctgactcctc aacaaaaatc cgggatgctg
     4861 cgacacaaca ttggccttga tgataaaaca ctccatggtg tgcaaactca acagcaatgt
     4921 ccaagtcggc atctgcaaac acaatgcaag ggctctttcc ccccagctcc agggtgactc
     4981 tcttcagatt gcttttccct gcagcttcct tgattaactt gccaacctgt gttgatccag
     5041 tgaaggccac cttgtcgaca tccatgtgag aggagatggc tgcccctgca gttggcccat
     5101 aaccagggac aatgtttacc acgccaggag gaaaccctgc ctcttttatt aaagatgcca
     5161 ggtgaagagc cgtgagagga gtttgctctg ctggcttgac aaccacggta ttcccacagc
     5221 taagggcagg gcctatcttc caaatgaaca tgagcattgg aaaattccag gggatgattt
     5281 ggccacacac tccaataggt tcacgtcttg tataagtgaa aatgtctcca tcacttggta
     5341 ttgtttgacc atgaatcttg tcagcccagc ctgcacagta ctttaatgct tttatgcagc
     5401 ctcctaaatc cgacaagtat gcattggcaa agactttccc accattgagt gcctccattg
     5461 tagccagcag cagacgatct ctctccatta agtcagccag cttgttcagc aggcggcccc
     5521 tctctgaagc atccatggtg cgccatggag agccaatctg gaaagcctgt cttgcagcct
     5581 tcacagcttt gtcaacatca gccttgtccc cttcttccac gtggcagatg acctcctcag
     5641 ttgcagggtt aagaactgga aatttcttgc cgctcactga attgtgccat tcattgttta
     5701 tgaagatctt ggtatgttga atcttcaagt cggccagtgg ggcagggact gcaggttgtg
     5761 caggcgaaga catggtggct agcataactt cgtataaagt atcctatacg aagttatttg
     5821 ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaata acttcgtata
     5881 atgtatgcta tacgaagtta tgaattcgat atcaagctta tcgataatca acctctggat
     5941 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     6001 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     6061 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     6121 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     6181 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     6241 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     6301 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     6361 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     6421 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     6481 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     6541 tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     6601 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     6661 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     6721 gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga
     6781 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     6841 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     6901 gcgca
//
