LOCUS       sequence-486663-        7483 bp    DNA     circular SYN 18-SEP-2025
DEFINITION  AAV vector expressing Mxra8 and tdTomato under cre-recombinase 
            control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7483)
  AUTHORS   Kim H, Xu C, Washington C, Lowmann M, Kebschull JM
  TITLE     Mapping the projectional architecture of the mouse midbrain 
            dopaminergic system using cell type-specific barcoding
  JOURNAL   bioRxiv 2025.06.25.661405; doi: 
            https://doi.org/10.1101/2025.06.25.661405
REFERENCE   2  (bases 1 to 7483)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.06.25.661405; doi: https://doi.org/10.1101/2025.06.25.661405"
COMMENT     Sequence Label: pAAV-CAG-FLEx-tdT-P2A-Mxra8
FEATURES             Location/Qualifiers
     source          1..7483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..131
                     /label=AAV2 ITR
     promoter        442..717
                     /label=chicken beta-actin promoter
     primer_bind     996..1018
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1080..1099
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1164..1197)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1208..1241)
                     /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)."
     CDS             complement(2536..2559)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2623..2679)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(3498..3521)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3696..3715)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3965..3983
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      4113..4122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4153..4186
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4197..4230
                     /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)."
     primer_bind     complement(4309..4329)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4842..4971
                     /label=AAV2 ITR
     rep_origin      complement(5208..5796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5288..5307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5967..6758,6759..6827))
                     /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     6590..6609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6828..6932)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6958..7413)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7095..7116)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7307..7326
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc
      181 taggaagagt accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      241 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      301 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      361 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      421 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      481 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      541 tgggggcggg gggggggggg gggcgcgcca ggcggggcgg ggcggggcga ggggcggggc
      601 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
      661 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      721 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
      781 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      841 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
      901 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcggggct gtccgcgggg
      961 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     1021 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     1081 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgctag cacgcgtgga
     1141 cggtaccctt caggtcgacg tcgataactt cgtatagcat acattatacg aagttatccg
     1201 aatcgcaata acttcgtata ggatacttta tacgaagtta tttggcgcgc cgctagctta
     1261 cttgcagtat tcttttcgaa actccttgtc caaatccaca tctttggcgg gcaatggtga
     1321 gtgagccaac tcagccctct ctttaagaat gttatttttg taatcaagtt gtatgtcctc
     1381 ggttctataa ggagcttgat cgcgagtggc aacggcgaat tcgaccatgt tgacatcctt
     1441 acccttactc ttcccggcct ttttgtcgga ggtcttacaa cccccagagt gtcgatatct
     1501 tgtagcaaga acaacagtta tcagaagcag gatgaaaagc agcagagtag cgagcacgta
     1561 cccgagctgc tgaaagaagt gagtatggtc ctcaggaaca atgacattta tgatagaatg
     1621 tcctctggtg agagtagggt ctggactggg tgctgagctg tgccctgatc cattacctgg
     1681 tgaggctctg gctggtggct caaaggctgg ctctgtgacc tggaggtgga agactctgcg
     1741 ctcatgaaga ccgcagtagt ggtggtggag atggcaacta tagatcccct cgtctgctcg
     1801 ttcgagttca tcgattctaa ggctgaagtc gcctcgtgca aaagcatttg tattgacact
     1861 taccctgtct cggaggaagg gaggtccgta agccctccgc tcgccacttg cgtagagatc
     1921 caaaagccta tcggctcgat cgtgtgagac gcctggcaac tgtctgtccc aatgtacaac
     1981 ttgttgggcc tcttccaaat ggcgatctgt ccaaacgtgg gctcgattga tacatgtcat
     2041 cagggctgga gcaccgtgtg cgactaccaa gacctccttt tcgccatccc aataagctcg
     2101 gctaaggagg ggatcttcgg taacttcaag gcggactgca agagattcgt caaggtggca
     2161 gtagtgatga tgcaggttac atgtataaac cccctcatct cctctatcca ctgcccttat
     2221 caagagagag aaatttccgt catggaatgc ggagggactg aggagcagcc gatcccggtc
     2281 tcgtggttcg tacacccgtt gttcccctgc actgtacata tcaacaagcc ttcgcctttg
     2341 gctcccagga ccacctgaca aatcccaatg gactaccctt tggcggtcgt gcaacctgtc
     2401 ttgggtccaa accattcggg gggattgaca gcgaaggact gcttgtgtac cggctgccaa
     2461 agaaaccaca ctttcggaga ccaatgaaga ggatgctgct gctgtcccgg aaggacctga
     2521 ggatcctcca tcgagcttgt catcgtcgtc cttgtagtcc atagcataca agccggtcag
     2581 tgagacaagc accagaaaga ctagggtcac cgagcgagcc attggcccag gattctcttc
     2641 gacatctcct gcttgtttca acagggagaa gttagtggct ccgcttccct tgtacagctc
     2701 gtccatgccg tacagaaaca gatggtgcct cccctcgctc cgctcatact gctccacaat
     2761 ggtgtagtct tcgttatggc tagtaatatc cagctttgtg tcgacgtaat agtagccagg
     2821 cagctgcacg ggtttcttgg ccatgtagat ggttttgaat tcgaccagat aatgccctcc
     2881 atctttcagc ttcagtgcct ggtggatctc tcccttcagc acgccgtctc tggggtacag
     2941 cctttctgtt gaagcctccc agcccattgt cttcttctgc atgactgggc catcaggggg
     3001 aaaattggtg ccgcgcattt tcaccttata gatcagagtg ccgtcctgca gagagctgtc
     3061 ctgggtgaca gtcaccaggc ccccgtcttc gaagttcatc actctttccc acttaaaccc
     3121 ctctgggaag ctcagtttct tgtaatctgg aatgtcagca gggtgtttca cgtaggcttt
     3181 acttccatac ataaactgag gagacaggat gtcccatgcg aatggcagag ggccgccctt
     3241 tgtcactttc agctttgctg tctgggtgcc ctcatagggc ctcccttccc cctctccttc
     3301 aatctcaaac tcgtgcccgt tcatgctgcc ctccatcctg actttaaacc tcataaactc
     3361 cttaatcacg gccatgttgt tgtcctcgga ggaggcggtg ccggagctgc cgctgccggt
     3421 gctgccggta ccatgcccca ggaacaggtg gtggcggccc tcggagcgct cgtactgttc
     3481 cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc ttggtgtcca cgtagtagta
     3541 gccgggcagt tgcacgggct tcttggccat gtagatggtc ttgaactcca ccaggtagtg
     3601 gccgccgtcc ttcagcttca gggcctggtg gatctcgccc ttcagcacgc cgtcgcgggg
     3661 gtacaggcgc tcggtggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc
     3721 gggggggaag ttggtgccgc gcatcttcac cttgtagatc agcgtgccgt cctgcaggga
     3781 ggagtcctgg gtcacggtca ccagaccgcc gtcctcgaag ttcatcacgc gctcccactt
     3841 gaagccctcg gggaaggaca gcttcttgta atcggggatg tcggcggggt gcttcacgta
     3901 cgccttggag ccgtacatga actgggggga caggatgtcc caggcgaagg gcagggggcc
     3961 gcccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc cctcgccctc
     4021 gccctcgatc tcgaactcgt ggccgttcat ggagccctcc atgcgcacct tgaagcgcat
     4081 gaactctttg atgacctcct cgcccttgct caccatggtg gcgctagcgg cgcgccaacc
     4141 tggccggcct tgataacttc gtataatgta tgctatacga agttatccga atcgcaataa
     4201 cttcgtataa agtatcctat acgaagttat cgaagatcat caagcttctc gagataatca
     4261 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4321 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4381 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     4441 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4501 gtttatttgt gaaatttgtg atgctattgc tttatttgta accatctagc tttatttgtg
     4561 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4621 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     4681 gcggccgcca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     4741 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc tagagcatgg
     4801 ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
     4861 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4921 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcattaa
     4981 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5041 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5401 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5641 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5761 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5881 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5941 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6001 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6061 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6121 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6181 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6241 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6301 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6361 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6421 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6481 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6541 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6601 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6661 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6721 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6781 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6841 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6901 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     6961 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     7021 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     7081 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     7141 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     7201 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     7261 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     7321 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     7381 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     7441 gttgggaagg gcgatcggtg cgggcctctt cgctattacg cca
//
