LOCUS       sequence-361933-        7283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV packaging plasmid, expressing Rep2 and AAV-cMG.WPP 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.WPP
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..36
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     28..45
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     29..45
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     55..74
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        55..73
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     102..118
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     103..119
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4129..4140
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4524..4544)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4524..4542)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4563..4579)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4563..4579)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4576..4598)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4587..4603
                     /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(4611..4617,4618..4635,4636..4641))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4656..4677
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4794..4811)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4965..5553)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5045..5064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5724..6515,6516..6584))
                     /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     6347..6366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6585..6689)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6715..7170)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6852..6873)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7064..7083
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
       61 gactcactat agggcgaatt gggtaccggg ccccccctcg atcgaggtcg acggtatcgg
      121 gggagctcgc agggtctcca ttttgaagcg ggaggtttga acgcgcagcc gccatgccgg
      181 ggttttacga gattgtgatt aaggtcccca gcgaccttga cgagcatctg cccggcattt
      241 ctgacagctt tgtgaactgg gtggccgaga aggaatggga gttgccgcca gattctgaca
      301 tggatctgaa tctgattgag caggcacccc tgaccgtggc cgagaagctg cagcgcgact
      361 ttctgacgga atggcgccgt gtgagtaagg ccccggaggc tcttttcttt gtgcaatttg
      421 agaagggaga gagctacttc cacatgcacg tgctcgtgga aaccaccggg gtgaaatcca
      481 tggttttggg acgtttcctg agtcagattc gcgaaaaact gattcagaga atttaccgcg
      541 ggatcgagcc gactttgcca aactggttcg cggtcacaaa gaccagaaat ggcgccggag
      601 gcgggaacaa ggtggtggat gagtgctaca tccccaatta cttgctcccc aaaacccagc
      661 ctgagctcca gtgggcgtgg actaatatgg aacagtattt aagcgcctgt ttgaatctca
      721 cggagcgtaa acggttggtg gcgcagcatc tgacgcacgt gtcgcagacg caggagcaga
      781 acaaagagaa tcagaatccc aattctgatg cgccggtgat cagatcaaaa acttcagcca
      841 ggtacatgga gctggtcggg tggctcgtgg acaaggggat tacctcggag aagcagtgga
      901 tccaggagga ccaggcctca tacatctcct tcaatgcggc ctccaactcg cggtcccaaa
      961 tcaaggctgc cttggacaat gcgggaaaga ttatgagcct gactaaaacc gcccccgact
     1021 acctggtggg ccagcagccc gtggaggaca tttccagcaa tcggatttat aaaattttgg
     1081 aactaaacgg gtacgatccc caatatgcgg cttccgtctt tctgggatgg gccacgaaaa
     1141 agttcggcaa gaggaacacc atctggctgt ttgggcctgc aactaccggg aagaccaaca
     1201 tcgcggaggc catagcccac actgtgccct tctacgggtg cgtaaactgg accaatgaga
     1261 actttccctt caacgactgt gtcgacaaga tggtgatctg gtgggaggag gggaagatga
     1321 ccgccaaggt cgtggagtcg gccaaagcca ttctcggagg aagcaaggtg cgcgtggacc
     1381 agaaatgcaa gtcctcggcc cagatagacc cgactcccgt gatcgtcacc tccaacacca
     1441 acatgtgcgc cgtgattgac gggaactcaa cgaccttcga acaccagcag ccgttgcaag
     1501 accggatgtt caaatttgaa ctcacccgcc gtctggatca tgactttggg aaggtcacca
     1561 agcaggaagt caaagacttt ttccggtggg caaaggatca cgtggttgag gtggagcatg
     1621 aattctacgt caaaaagggt ggagccaaga aaagacccgc ccccagtgac gcagatataa
     1681 gtgagcccaa acgggtgcgc gagtcagttg cgcagccatc gacgtcagac gcggaagctt
     1741 cgatcaacta cgcagacagg taccaaaaca aatgttctcg tcacgtgggc atgaatctga
     1801 tgctgtttcc ctgcagacaa tgcgagagaa tgaatcagaa ttcaaatatc tgcttcactc
     1861 acggacagaa agactgttta gagtgctttc ccgtgtcaga atctcaaccc gtttctgtcg
     1921 tcaaaaaggc gtatcagaaa ctgtgctaca ttcatcatat catgggaaag gtgccagacg
     1981 cttgcactgc ctgcgatctg gtcaatgtgg atttggatga ctgcatcttt gaacaataaa
     2041 tgatttaaat caggtatggc tgccgatggt tatcttccag attggctcga ggacaacctt
     2101 agtgaaggaa ttcgcgagtg gtgggctttg aaacctggag cccctcaacc caaggcaaat
     2161 caacaacatc aagacaacgc tcgaggtctt gtgcttccgg gttacaaata ccttggaccc
     2221 ggcaacggac tcgacaaggg ggagccggtc aacgcagcag acgcggcggc cctcgagcac
     2281 gacaaggcct acgaccagca gctcaaggcc ggagacaacc cgtacctcaa gtacaaccac
     2341 gccgacgccg agttccagga gcggctcaaa gaagatacgt cttttggggg caacctcggg
     2401 cgagcagtct tccaggccaa aaagaggctt cttgaacctc ttggtctggt tgaggaagcg
     2461 gctaagacgg ctcctggaaa gaagaggcct gtagagcagt ctcctcagga accggactcc
     2521 tccgcgggta ttggcaaatc gggtgcacag cccgctaaaa agagactcaa tttcggtcag
     2581 actggcgaca cagagtcagt cccagaccct caaccaatcg gagaacctcc cgcagccccc
     2641 tcaggtgtgg gatctcttac aatggcttca ggtggtggcg caccagtggc agacaataac
     2701 gaaggtgccg atggagtggg tagttcctcg ggaaattggc attgcgattc ccaatggctg
     2761 ggggacagag tcatcaccac cagcacccga acctgggccc tgcccaccta caacaatcac
     2821 ctctacaagc aaatctccaa cagcacatct ggaggatctt caaatgacaa cgcctacttc
     2881 ggctacagca ccccctgggg gtattttgac ttcaacagat tccactgcca cttctcacca
     2941 cgtgactggc agcgactcat caacaacaac tggggattcc ggcctaagcg actcaacttc
     3001 aagctcttca acattcaggt caaagaggtt acggacaaca atggagtcaa gaccatcgcc
     3061 aataacctta ccagcacggt ccaggtcttc acggactcag actatcagct cccgtacgtg
     3121 ctcgggtcgg ctcacgaggg ctgcctcccg ccgttcccag cggacgtttt catgattcct
     3181 cagtacgggt atctgacgct taatgatgga agccaggccg tgggtcgttc gtccttttac
     3241 tgcctggaat atttcccgtc gcaaatgcta agaacgggta acaacttcca gttcagctac
     3301 gagtttgaga acgtaccttt ccatagcagc tacgctcaca gccaaagcct ggaccgacta
     3361 atgaatccac tcatcgacca atacttgtac tatctctcta gaactattaa cggttctgga
     3421 cagaatcaac aaacgctaaa attcagtgtg gccggaccca gcaacatggc tgtccaggga
     3481 agaaactaca tacctggacc cagctaccga caacaacgtg tctcaaccac tgtgactcaa
     3541 aacaacaaca gcgaatttgc ttggcctgga gcttcttctt gggctctcaa tggacgtaat
     3601 agcttgatga atcctggacc tgctatggcc agccacaaag aaggagagga ccgtttcttt
     3661 cctttgtctg gatctttaat ttttggcaaa caaggtaccg gtagagacaa cgtggatgcg
     3721 gacaaagtca tgataaccaa cgaagaagaa attaaaacta ctaacccggt agcaacggag
     3781 tcctatggac aagtggccac aaaccaccag agtgcccaat ggccgccgaa gactacgtct
     3841 gcacaggcgc agaccggttg ggttcaaaac caaggaatac ttccgggtat ggtttggcag
     3901 gacagagatg tgtacctgca aggacccatt tgggccaaaa ttcctcacac ggacggcaac
     3961 tttcaccctt ctccgctgat gggagggttt ggaatgaagc acccgcctcc tcagatcctc
     4021 atcaaaaaca cacctgtacc tgcggatcct ccaacggcct tcaacaagga caagctgaac
     4081 tctttcatca cccagtattc tactggccaa gtcagcgtgg agatcgagtg ggagctgcag
     4141 aaggaaaaca gcaagcgctg gaacccggag atccagtaca cttccaacta ttacaagtct
     4201 aataatgttg aatttgctgt taatactgaa ggtgtatata gtgaaccccg ccccattggc
     4261 accagatacc tgactcgtaa tctgtaagtt taaacctgca ggactagagg tcctgtatta
     4321 gaggtcacgt gagtgttttg cgacattttg cgacaccatg tggtcacgct gggtatttaa
     4381 gcccgagtga gcacgcaggg tctccatttt gaagcgggag gtttgaacgc gcagccgcca
     4441 agccgaattc tgcagatatc catcacactg gcggccgctc gactagagcg gccgccaccg
     4501 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     4561 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4621 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4681 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4741 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4801 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4861 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4921 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4981 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5041 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5101 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5161 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5221 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5281 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5341 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5401 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5461 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5521 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5581 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5641 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5701 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5761 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5821 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5881 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5941 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6001 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6061 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6121 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6181 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6241 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6301 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6361 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6421 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6481 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6541 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6601 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6661 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     6721 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     6781 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     6841 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     6901 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     6961 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     7021 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     7081 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     7141 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     7201 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     7261 ctgcaaggcg attaagttgg gta
//
