LOCUS       sequence-361936-        7283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV packaging plasmid, expressing Rep2 and AAV-cMG capsid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7283)
  AUTHORS   Lin R, Zhou Y, Yan T, Wang R, Li H, Wu Z, Zhang X, Zhou X, Zhao F, 
            Zhang L, Li Y, Luo M
  TITLE     Directed evolution of adeno-associated virus for efficient gene 
            delivery to microglia.
  JOURNAL   Nat Methods. 2022 Jul 25. pii: 10.1038/s41592-022-01547-7. doi: 
            10.1038/s41592-022-01547-7.
  PUBMED    35879607
REFERENCE   2  (bases 1 to 7283)
  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: "Nat 
            Methods. 2022 Jul 25. pii: 10.1038/s41592-022-01547-7. doi: 
            10.1038/s41592-022-01547-7."
COMMENT     Sequence Label: rAAV2/cMG
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     64..83
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        64..82
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     111..127
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     112..128
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4138..4149
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4533..4553)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4533..4551)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4572..4588)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4572..4588)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4585..4607)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4596..4612
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4620..4626,4627..4644,4645..4650))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4665..4686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4803..4820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4974..5562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5054..5073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5733..6524,6525..6593))
                     /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     6356..6375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6594..6698)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6724..7179)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6861..6882)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7073..7092
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
       61 gcgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga tcgaggtcga
      121 cggtatcggg ggagctcgca gggtctccat tttgaagcgg gaggtttgaa cgcgcagccg
      181 ccatgccggg gttttacgag attgtgatta aggtccccag cgaccttgac gagcatctgc
      241 ccggcatttc tgacagcttt gtgaactggg tggccgagaa ggaatgggag ttgccgccag
      301 attctgacat ggatctgaat ctgattgagc aggcacccct gaccgtggcc gagaagctgc
      361 agcgcgactt tctgacggaa tggcgccgtg tgagtaaggc cccggaggct cttttctttg
      421 tgcaatttga gaagggagag agctacttcc acatgcacgt gctcgtggaa accaccgggg
      481 tgaaatccat ggttttggga cgtttcctga gtcagattcg cgaaaaactg attcagagaa
      541 tttaccgcgg gatcgagccg actttgccaa actggttcgc ggtcacaaag accagaaatg
      601 gcgccggagg cgggaacaag gtggtggatg agtgctacat ccccaattac ttgctcccca
      661 aaacccagcc tgagctccag tgggcgtgga ctaatatgga acagtattta agcgcctgtt
      721 tgaatctcac ggagcgtaaa cggttggtgg cgcagcatct gacgcacgtg tcgcagacgc
      781 aggagcagaa caaagagaat cagaatccca attctgatgc gccggtgatc agatcaaaaa
      841 cttcagccag gtacatggag ctggtcgggt ggctcgtgga caaggggatt acctcggaga
      901 agcagtggat ccaggaggac caggcctcat acatctcctt caatgcggcc tccaactcgc
      961 ggtcccaaat caaggctgcc ttggacaatg cgggaaagat tatgagcctg actaaaaccg
     1021 cccccgacta cctggtgggc cagcagcccg tggaggacat ttccagcaat cggatttata
     1081 aaattttgga actaaacggg tacgatcccc aatatgcggc ttccgtcttt ctgggatggg
     1141 ccacgaaaaa gttcggcaag aggaacacca tctggctgtt tgggcctgca actaccggga
     1201 agaccaacat cgcggaggcc atagcccaca ctgtgccctt ctacgggtgc gtaaactgga
     1261 ccaatgagaa ctttcccttc aacgactgtg tcgacaagat ggtgatctgg tgggaggagg
     1321 ggaagatgac cgccaaggtc gtggagtcgg ccaaagccat tctcggagga agcaaggtgc
     1381 gcgtggacca gaaatgcaag tcctcggccc agatagaccc gactcccgtg atcgtcacct
     1441 ccaacaccaa catgtgcgcc gtgattgacg ggaactcaac gaccttcgaa caccagcagc
     1501 cgttgcaaga ccggatgttc aaatttgaac tcacccgccg tctggatcat gactttggga
     1561 aggtcaccaa gcaggaagtc aaagactttt tccggtgggc aaaggatcac gtggttgagg
     1621 tggagcatga attctacgtc aaaaagggtg gagccaagaa aagacccgcc cccagtgacg
     1681 cagatataag tgagcccaaa cgggtgcgcg agtcagttgc gcagccatcg acgtcagacg
     1741 cggaagcttc gatcaactac gcagacaggt accaaaacaa atgttctcgt cacgtgggca
     1801 tgaatctgat gctgtttccc tgcagacaat gcgagagaat gaatcagaat tcaaatatct
     1861 gcttcactca cggacagaaa gactgtttag agtgctttcc cgtgtcagaa tctcaacccg
     1921 tttctgtcgt caaaaaggcg tatcagaaac tgtgctacat tcatcatatc atgggaaagg
     1981 tgccagacgc ttgcactgcc tgcgatctgg tcaatgtgga tttggatgac tgcatctttg
     2041 aacaataaat gatttaaatc aggtatggct gccgatggtt atcttccaga ttggctcgag
     2101 gacaacctta gtgaaggaat tcgcgagtgg tgggctttga aacctggagc ccctcaaccc
     2161 aaggcaaatc aacaacatca agacaacgct cgaggtcttg tgcttccggg ttacaaatac
     2221 cttggacccg gcaacggact cgacaagggg gagccggtca acgcagcaga cgcggcggcc
     2281 ctcgagcacg acaaggccta cgaccagcag ctcaaggccg gagacaaccc gtacctcaag
     2341 tacaaccacg ccgacgccga gttccaggag cggctcaaag aagatacgtc ttttgggggc
     2401 aacctcgggc gagcagtctt ccaggccaaa aagaggcttc ttgaacctct tggtctggtt
     2461 gaggaagcgg ctaagacggc tcctggaaag aagaggcctg tagagcagtc tcctcaggaa
     2521 ccggactcct ccgcgggtat tggcaaatcg ggtgcacagc ccgctaaaaa gagactcaat
     2581 ttcggtcaga ctggcgacac agagtcagtc ccagaccctc aaccaatcgg agaacctccc
     2641 gcagccccct caggtgtggg atctcttaca atggcttcag gtggtggcgc accagtggca
     2701 gacaataacg aaggtgccga tggagtgggt agttcctcgg gaaattggca ttgcgattcc
     2761 caatggctgg gggacagagt catcaccacc agcacccgaa cctgggccct gcccacctac
     2821 aacaatcacc tctacaagca aatctccaac agcacatctg gaggatcttc aaatgacaac
     2881 gcctacttcg gctacagcac cccctggggg tattttgact tcaacagatt ccactgccac
     2941 ttctcaccac gtgactggca gcgactcatc aacaacaact ggggattccg gcctaagcga
     3001 ctcaacttca agctcttcaa cattcaggtc aaagaggtta cggacaacaa tggagtcaag
     3061 accatcgcca ataaccttac cagcacggtc caggtcttca cggactcaga ctatcagctc
     3121 ccgtacgtgc tcgggtcggc tcacgagggc tgcctcccgc cgttcccagc ggacgttttc
     3181 atgattcctc agtacgggta tctgacgctt aatgatggaa gccaggccgt gggtcgttcg
     3241 tccttttact gcctggaata tttcccgtcg caaatgctaa gaacgggtaa caacttccag
     3301 ttcagctacg agtttgagaa cgtacctttc catagcagct acgctcacag ccaaagcctg
     3361 gaccgactaa tgaatccact catcgaccaa tacttgtact atctctctag aactattaac
     3421 ggttctggac agaatcaaca aacgctaaaa ttcagtgtgg ccggacccag caacatggct
     3481 gtccagggaa gaaactacat acctggaccc agctaccgac aacaacgtgt ctcaaccact
     3541 gtgactcaaa acaacaacag cgaatttgct tggcctggag cttcttcttg ggctctcaat
     3601 ggacgtaata gcttgatgaa tcctggacct gctatggcca gccacaaaga aggagaggac
     3661 cgtttctttc ctttgtctgg atctttaatt tttggcaaac aaggtaccgg tagagacaac
     3721 gtggatgcgg acaaagtcat gataaccaac gaagaagaaa ttaaaactac taacccggta
     3781 gcaacggagt cctatggaca agtggccaca aaccaccaga gtgcccaaca gaggcctccg
     3841 cgtccggctg atcaggcgca gaccggttgg gttcaaaacc aaggaatact tccgggtatg
     3901 gtttggcagg acagagatgt gtacctgcaa ggacccattt gggccaaaat tcctcacacg
     3961 gacggcaact ttcacccttc tccgctgatg ggagggtttg gaatgaagca cccgcctcct
     4021 cagatcctca tcaaaaacac acctgtacct gcggatcctc caacggcctt caacaaggac
     4081 aagctgaact ctttcatcac ccagtattct actggccaag tcagcgtgga gatcgagtgg
     4141 gagctgcaga aggaaaacag caagcgctgg aacccggaga tccagtacac ttccaactat
     4201 tacaagtcta ataatgttga atttgctgtt aatactgaag gtgtatatag tgaaccccgc
     4261 cccattggca ccagatacct gactcgtaat ctgtaagttt aaacctgcag gactagaggt
     4321 cctgtattag aggtcacgtg agtgttttgc gacattttgc gacaccatgt ggtcacgctg
     4381 ggtatttaag cccgagtgag cacgcagggt ctccattttg aagcgggagg tttgaacgcg
     4441 cagccgccaa gccgaattct gcagatatcc atcacactgg cggccgctcg actagagcgg
     4501 ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     4561 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4621 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4681 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4741 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4801 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4861 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5401 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5701 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5761 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5821 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5881 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5941 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6001 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6061 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6121 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6181 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6241 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6301 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6361 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6421 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6481 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6541 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6601 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6661 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6721 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6781 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     6841 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     6901 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6961 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     7021 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     7081 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     7141 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     7201 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     7261 ggggatgtgc tgcaaggcga tta
//
