LOCUS       sequence-486662-        7560 bp    DNA     circular SYN 18-SEP-2025
DEFINITION  AAV vector expressing Mxra8 and tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7560)
  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 7560)
  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-tdT-P2A-Mxra8
FEATURES             Location/Qualifiers
     source          1..7560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..131
                     /label=AAV2 ITR
     promoter        442..715
                     /label=chicken beta-actin promoter
     primer_bind     994..1016
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1078..1097
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2571..2627
                     /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"
     CDS             2691..2714
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4005..4593
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4058..4078)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4476..4487)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4636..4757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4647..4666)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4701..4720
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4919..5048
                     /label=AAV2 ITR
     rep_origin      complement(5285..5873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5365..5384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6044..6835,6836..6904))
                     /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     6667..6686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6905..7009)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7035..7490)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7172..7193)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7384..7403
                     /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 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
      601 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
      661 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
      721 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
      781 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      841 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
      901 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggctgt ccgcgggggg
      961 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1021 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1081 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgctagcc accatggtga
     1141 gtaagggcga ggaagtgatc aaagagttca tgcggtttaa ggtgagaatg gaaggaagca
     1201 tgaacggcca cgagttcgaa attgagggag aaggagaggg acggccctac gagggcaccc
     1261 agacagccaa gctgaaagtg acaaagggcg ggcctctgcc attcgcttgg gacatcctga
     1321 gcccacagtt tatgtacggc tccaaggcct atgtgaaaca tccagctgac attcccgatt
     1381 ataagaaact gagcttcccc gaggggttta agtgggaaag agtgatgaac ttcgaggacg
     1441 gaggcctggt gactgtgacc caggacagct ccctgcagga tgggaccctg atctacaagg
     1501 tgaaaatgag agggacaaat tttccccctg atggacctgt gatgcagaag aaaactatgg
     1561 gatgggaggc ctccaccgaa aggctgtatc cacgcgacgg ggtgctgaaa ggagaaatcc
     1621 accaggctct gaagctgaaa gatgggggac attacctggt ggagttcaag acaatctaca
     1681 tggccaagaa acctgtgcag ctgccaggct actattacgt ggacacaaaa ctggatatca
     1741 cttcacacaa cgaggactac actattgtgg agcagtatga acggagcgag gggagacacc
     1801 atctgttcct gggccatggg actggaagta ccggctcagg gtctagtgga accgcctcaa
     1861 gcgaggataa caatatggct gtgatcaaag agttcatgag gtttaaggtg cgcatggagg
     1921 gcagcatgaa tgggcacgaa tttgagattg aaggagaggg cgaagggagg ccttacgagg
     1981 gcacacagac tgccaagctg aaagtgacca agggaggacc actgcctttc gcttgggata
     2041 tcctgtctcc tcagtttatg tacggaagta aggcctatgt caagcatccc gctgacattc
     2101 ctgattacaa gaaactgtct ttcccagagg gctttaagtg ggagagagtg atgaattttg
     2161 aagatggagg cctggtgacc gtgacacagg actcctctct gcaggatggc actctgatct
     2221 acaaagtcaa aatgcgcggc accaattttc cacccgatgg gcccgtgatg cagaagaaaa
     2281 caatggggtg ggaggccagc actgaacggc tgtatcctag agacggagtg ctgaagggcg
     2341 aaatccacca ggccctgaag ctgaaagacg gcggccacta cctggtggag ttcaaaacca
     2401 tctacatggc caagaaacca gtgcagctgc ccggctatta ctatgtggac accaagctgg
     2461 atatcacatc ccacaatgaa gactacacca ttgtggaaca gtatgagagg tctgaaggac
     2521 gccaccatct gtttctgtac ggcatggatg agctgtataa gggaagcgga gccactaact
     2581 tctccctgtt gaaacaagca ggagatgtcg aagagaatcc tgggccaatg gctcgctcgg
     2641 tgaccctagt ctttctggtg cttgtctcac tgaccggctt gtatgctatg gactacaagg
     2701 acgacgatga caagctcgat ggaggatcct caggtccttc cgggacagca gcagcatcct
     2761 cttcattggt ctccgaaagt gtggtttctt tggcagccgg tacacaagca gtccttcgct
     2821 gtcaatcccc ccgaatggtt tggacccaag acaggttgca cgaccgccaa agggtagtcc
     2881 attgggattt gtcaggtggt cctgggagcc aaaggcgaag gcttgttgat atgtacagtg
     2941 caggggaaca acgggtgtac gaaccacgag accgggatcg gctgctcctc agtccctccg
     3001 cattccatga cggaaatttc tctctcttga taagggcagt ggatagagga gatgaggggg
     3061 tttatacatg taacctgcat catcactact gccaccttga cgaatctctt gcagtccgcc
     3121 ttgaagttac cgaagatccc ctccttagcc gagcttattg ggatggcgaa aaggaggtct
     3181 tggtagtcgc acacggtgct ccagccctga tgacatgtat caatcgagcc cacgtttgga
     3241 cagatcgcca tttggaagag gcccaacaag ttgtacattg ggacagacag ttgccaggcg
     3301 tctcacacga tcgagccgat aggcttttgg atctctacgc aagtggcgag cggagggctt
     3361 acggacctcc cttcctccga gacagggtaa gtgtcaatac aaatgctttt gcacgaggcg
     3421 acttcagcct tagaatcgat gaactcgaac gagcagacga ggggatctat agttgccatc
     3481 tccaccacca ctactgcggt cttcatgagc gcagagtctt ccacctccag gtcacagagc
     3541 cagcctttga gccaccagcc agagcctcac caggtaatgg atcagggcac agctcagcac
     3601 ccagtccaga ccctactctc accagaggac attctatcat aaatgtcatt gttcctgagg
     3661 accatactca cttctttcag cagctcgggt acgtgctcgc tactctgctg cttttcatcc
     3721 tgcttctgat aactgttgtt cttgctacaa gatatcgaca ctctgggggt tgtaagacct
     3781 ccgacaaaaa ggccgggaag agtaagggta aggatgtcaa catggtcgaa ttcgccgttg
     3841 ccactcgcga tcaagctcct tatagaaccg aggacataca acttgattac aaaaataaca
     3901 ttcttaaaga gagggctgag ttggctcact caccattgcc cgccaaagat gtggatttgg
     3961 acaaggagtt tcgaaaagaa tactgcaagt aaaagcttat cgataatcaa cctctggatt
     4021 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     4081 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     4141 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     4201 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     4261 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     4321 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4381 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     4441 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4501 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     4561 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc
     4621 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     4681 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     4741 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     4801 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     4861 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     4921 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     4981 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     5041 gcgcgcagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7021 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     7081 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     7141 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     7201 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     7261 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     7321 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     7381 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     7441 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     7501 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
//
