LOCUS       sequence-361941-        7283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV packaging plasmid, expressing Rep2 and AAV-MG1.2 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/MG1.2
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..33
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     25..42
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     26..42
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     52..71
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        52..70
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     99..115
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     100..116
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4126..4137
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4521..4541)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4521..4539)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4560..4576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4560..4576)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4573..4595)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4584..4600
                     /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(4608..4614,4615..4632,4633..4638))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4653..4674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4791..4808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4962..5550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5042..5061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5721..6512,6513..6581))
                     /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     6344..6363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6582..6686)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6712..7167)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6849..6870)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7061..7080
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
       61 tcactatagg gcgaattggg taccgggccc cccctcgatc gaggtcgacg gtatcggggg
      121 agctcgcagg gtctccattt tgaagcggga ggtttgaacg cgcagccgcc atgccggggt
      181 tttacgagat tgtgattaag gtccccagcg accttgacga gcatctgccc ggcatttctg
      241 acagctttgt gaactgggtg gccgagaagg aatgggagtt gccgccagat tctgacatgg
      301 atctgaatct gattgagcag gcacccctga ccgtggccga gaagctgcag cgcgactttc
      361 tgacggaatg gcgccgtgtg agtaaggccc cggaggctct tttctttgtg caatttgaga
      421 agggagagag ctacttccac atgcacgtgc tcgtggaaac caccggggtg aaatccatgg
      481 ttttgggacg tttcctgagt cagattcgcg aaaaactgat tcagagaatt taccgcggga
      541 tcgagccgac tttgccaaac tggttcgcgg tcacaaagac cagaaatggc gccggaggcg
      601 ggaacaaggt ggtggatgag tgctacatcc ccaattactt gctccccaaa acccagcctg
      661 agctccagtg ggcgtggact aatatggaac agtatttaag cgcctgtttg aatctcacgg
      721 agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc gcagacgcag gagcagaaca
      781 aagagaatca gaatcccaat tctgatgcgc cggtgatcag atcaaaaact tcagccaggt
      841 acatggagct ggtcgggtgg ctcgtggaca aggggattac ctcggagaag cagtggatcc
      901 aggaggacca ggcctcatac atctccttca atgcggcctc caactcgcgg tcccaaatca
      961 aggctgcctt ggacaatgcg ggaaagatta tgagcctgac taaaaccgcc cccgactacc
     1021 tggtgggcca gcagcccgtg gaggacattt ccagcaatcg gatttataaa attttggaac
     1081 taaacgggta cgatccccaa tatgcggctt ccgtctttct gggatgggcc acgaaaaagt
     1141 tcggcaagag gaacaccatc tggctgtttg ggcctgcaac taccgggaag accaacatcg
     1201 cggaggccat agcccacact gtgcccttct acgggtgcgt aaactggacc aatgagaact
     1261 ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg ggaggagggg aagatgaccg
     1321 ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag caaggtgcgc gtggaccaga
     1381 aatgcaagtc ctcggcccag atagacccga ctcccgtgat cgtcacctcc aacaccaaca
     1441 tgtgcgccgt gattgacggg aactcaacga ccttcgaaca ccagcagccg ttgcaagacc
     1501 ggatgttcaa atttgaactc acccgccgtc tggatcatga ctttgggaag gtcaccaagc
     1561 aggaagtcaa agactttttc cggtgggcaa aggatcacgt ggttgaggtg gagcatgaat
     1621 tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc cagtgacgca gatataagtg
     1681 agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac gtcagacgcg gaagcttcga
     1741 tcaactacgc agacaggtac caaaacaaat gttctcgtca cgtgggcatg aatctgatgc
     1801 tgtttccctg cagacaatgc gagagaatga atcagaattc aaatatctgc ttcactcacg
     1861 gacagaaaga ctgtttagag tgctttcccg tgtcagaatc tcaacccgtt tctgtcgtca
     1921 aaaaggcgta tcagaaactg tgctacattc atcatatcat gggaaaggtg ccagacgctt
     1981 gcactgcctg cgatctggtc aatgtggatt tggatgactg catctttgaa caataaatga
     2041 tttaaatcag gtatggctgc cgatggttat cttccagatt ggctcgagga caaccttagt
     2101 gaaggaattc gcgagtggtg ggctttgaaa cctggagccc ctcaacccaa ggcaaatcaa
     2161 caacatcaag acaacgctcg aggtcttgtg cttccgggtt acaaatacct tggacccggc
     2221 aacggactcg acaaggggga gccggtcaac gcagcagacg cggcggccct cgagcacgac
     2281 aaggcctacg accagcagct caaggccgga gacaacccgt acctcaagta caaccacgcc
     2341 gacgccgagt tccaggagcg gctcaaagaa gatacgtctt ttgggggcaa cctcgggcga
     2401 gcagtcttcc aggccaaaaa gaggcttctt gaacctcttg gtctggttga ggaagcggct
     2461 aagacggctc ctggaaagaa gaggcctgta gagcagtctc ctcaggaacc ggactcctcc
     2521 gcgggtattg gcaaatcggg tgcacagccc gctaaaaaga gactcaattt cggtcagact
     2581 ggcgacacag agtcagtccc agaccctcaa ccaatcggag aacctcccgc agccccctca
     2641 ggtgtgggat ctcttacaat ggcttcaggt ggtggcgcac cagtggcaga caataacgaa
     2701 ggtgccgatg gagtgggtag ttcctcggga aattggcatt gcgattccca atggctgggg
     2761 gacagagtca tcaccaccag cacccgaacc tgggccctgc ccacctacaa caatcacctc
     2821 tacaagcaaa tctccaacag cacatctgga ggatcttcaa atgacaacgc ctacttcggc
     2881 tacagcaccc cctgggggta ttttgacttc aacagattcc actgccactt ctcaccacgt
     2941 gactggcagc gactcatcaa caacaactgg ggattccggc ctaagcgact caacttcaag
     3001 ctcttcaaca ttcaggtcaa agaggttacg gacaacaatg gagtcaagac catcgccaat
     3061 aaccttacca gcacggtcca ggtcttcacg gactcagact atcagctccc gtacgtgctc
     3121 gggtcggctc acgagggctg cctcccgccg ttcccagcgg acgttttcat gattcctcag
     3181 tacgggtatc tgacgcttaa tgatggaagc caggccgtgg gtcgttcgtc cttttactgc
     3241 ctggaatatt tcccgtcgca aatgctaaga acgggtaaca acttccagtt cagctacgag
     3301 tttgagaacg tacctttcca tagcagctac gctcacagcc aaagcctgga ccgactaatg
     3361 aatccactca tcgaccaata cttgtactat ctctctagaa ctattaacgg ttctggacag
     3421 aatcaacaaa cgctaaaatt cagtgtggcc ggacccagca acatggctgt ccagggaaga
     3481 aactacatac ctggacccag ctaccgacaa caacgtgtct caaccactgt gactcaaaac
     3541 aacaacagcg aatttgcttg gcctggagct tcttcttggg ctctcaatgg acgtaatagc
     3601 ttgatgaatc ctggacctgc tatggccagc cacaaagaag gagaggaccg tttctttcct
     3661 ttgtctggat ctttaatttt tggcaaacaa ggtaccggta gagacaacgt ggatgcggac
     3721 aaagtcatga taaccaacga agaagaaatt aaaactacta acccggtagc aacggagtcc
     3781 tatggacaag tggccacaaa ccaccagagt gccactgagc ctccgaagac tacgtctgca
     3841 caggcgcaga ccggttgggt tcaaaaccaa ggaatacttc cgggtatggt ttggcaggac
     3901 agagatgtgt acctgcaagg acccatttgg gccaaaattc ctcacacgga cggcaacttt
     3961 cacccttctc cgctgatggg agggtttgga atgaagcacc cgcctcctca gatcctcatc
     4021 aaaaacacac ctgtacctgc ggatcctcca acggccttca acaaggacaa gctgaactct
     4081 ttcatcaccc agtattctac tggccaagtc agcgtggaga tcgagtggga gctgcagaag
     4141 gaaaacagca agcgctggaa cccggagatc cagtacactt ccaactatta caagtctaat
     4201 aatgttgaat ttgctgttaa tactgaaggt gtatatagtg aaccccgccc cattggcacc
     4261 agatacctga ctcgtaatct gtaagtttaa acctgcagga ctagaggtcc tgtattagag
     4321 gtcacgtgag tgttttgcga cattttgcga caccatgtgg tcacgctggg tatttaagcc
     4381 cgagtgagca cgcagggtct ccattttgaa gcgggaggtt tgaacgcgca gccgccaagc
     4441 cgaattctgc agatatccat cacactggcg gccgctcgac tagagcggcc gccaccgcgg
     4501 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     4561 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4621 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4681 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4741 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4801 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4861 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5401 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5581 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5641 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5701 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5761 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5821 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5881 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5941 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6001 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6061 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6121 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6181 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6241 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6301 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6361 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6421 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6481 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6541 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6601 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6661 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     6721 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     6781 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     6841 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     6901 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     6961 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     7021 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     7081 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     7141 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     7201 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     7261 caaggcgatt aagttgggta acg
//
