LOCUS       sequence-361939-        7283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV packaging plasmid, expressing Rep2 and AAV-cMG.QRP 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.QRP
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..68
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     60..77
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     61..77
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     87..106
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        87..105
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     134..150
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     135..151
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4161..4172
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4556..4576)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4556..4574)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4595..4611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4595..4611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4608..4630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4619..4635
                     /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(4643..4649,4650..4667,4668..4673))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4688..4709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4826..4843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4997..5585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5077..5096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5756..6547,6548..6616))
                     /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     6379..6398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6617..6721)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6747..7202)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6884..6905)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7096..7115
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
       61 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
      121 ggccccccct cgatcgaggt cgacggtatc gggggagctc gcagggtctc cattttgaag
      181 cgggaggttt gaacgcgcag ccgccatgcc ggggttttac gagattgtga ttaaggtccc
      241 cagcgacctt gacgagcatc tgcccggcat ttctgacagc tttgtgaact gggtggccga
      301 gaaggaatgg gagttgccgc cagattctga catggatctg aatctgattg agcaggcacc
      361 cctgaccgtg gccgagaagc tgcagcgcga ctttctgacg gaatggcgcc gtgtgagtaa
      421 ggccccggag gctcttttct ttgtgcaatt tgagaaggga gagagctact tccacatgca
      481 cgtgctcgtg gaaaccaccg gggtgaaatc catggttttg ggacgtttcc tgagtcagat
      541 tcgcgaaaaa ctgattcaga gaatttaccg cgggatcgag ccgactttgc caaactggtt
      601 cgcggtcaca aagaccagaa atggcgccgg aggcgggaac aaggtggtgg atgagtgcta
      661 catccccaat tacttgctcc ccaaaaccca gcctgagctc cagtgggcgt ggactaatat
      721 ggaacagtat ttaagcgcct gtttgaatct cacggagcgt aaacggttgg tggcgcagca
      781 tctgacgcac gtgtcgcaga cgcaggagca gaacaaagag aatcagaatc ccaattctga
      841 tgcgccggtg atcagatcaa aaacttcagc caggtacatg gagctggtcg ggtggctcgt
      901 ggacaagggg attacctcgg agaagcagtg gatccaggag gaccaggcct catacatctc
      961 cttcaatgcg gcctccaact cgcggtccca aatcaaggct gccttggaca atgcgggaaa
     1021 gattatgagc ctgactaaaa ccgcccccga ctacctggtg ggccagcagc ccgtggagga
     1081 catttccagc aatcggattt ataaaatttt ggaactaaac gggtacgatc cccaatatgc
     1141 ggcttccgtc tttctgggat gggccacgaa aaagttcggc aagaggaaca ccatctggct
     1201 gtttgggcct gcaactaccg ggaagaccaa catcgcggag gccatagccc acactgtgcc
     1261 cttctacggg tgcgtaaact ggaccaatga gaactttccc ttcaacgact gtgtcgacaa
     1321 gatggtgatc tggtgggagg aggggaagat gaccgccaag gtcgtggagt cggccaaagc
     1381 cattctcgga ggaagcaagg tgcgcgtgga ccagaaatgc aagtcctcgg cccagataga
     1441 cccgactccc gtgatcgtca cctccaacac caacatgtgc gccgtgattg acgggaactc
     1501 aacgaccttc gaacaccagc agccgttgca agaccggatg ttcaaatttg aactcacccg
     1561 ccgtctggat catgactttg ggaaggtcac caagcaggaa gtcaaagact ttttccggtg
     1621 ggcaaaggat cacgtggttg aggtggagca tgaattctac gtcaaaaagg gtggagccaa
     1681 gaaaagaccc gcccccagtg acgcagatat aagtgagccc aaacgggtgc gcgagtcagt
     1741 tgcgcagcca tcgacgtcag acgcggaagc ttcgatcaac tacgcagaca ggtaccaaaa
     1801 caaatgttct cgtcacgtgg gcatgaatct gatgctgttt ccctgcagac aatgcgagag
     1861 aatgaatcag aattcaaata tctgcttcac tcacggacag aaagactgtt tagagtgctt
     1921 tcccgtgtca gaatctcaac ccgtttctgt cgtcaaaaag gcgtatcaga aactgtgcta
     1981 cattcatcat atcatgggaa aggtgccaga cgcttgcact gcctgcgatc tggtcaatgt
     2041 ggatttggat gactgcatct ttgaacaata aatgatttaa atcaggtatg gctgccgatg
     2101 gttatcttcc agattggctc gaggacaacc ttagtgaagg aattcgcgag tggtgggctt
     2161 tgaaacctgg agcccctcaa cccaaggcaa atcaacaaca tcaagacaac gctcgaggtc
     2221 ttgtgcttcc gggttacaaa taccttggac ccggcaacgg actcgacaag ggggagccgg
     2281 tcaacgcagc agacgcggcg gccctcgagc acgacaaggc ctacgaccag cagctcaagg
     2341 ccggagacaa cccgtacctc aagtacaacc acgccgacgc cgagttccag gagcggctca
     2401 aagaagatac gtcttttggg ggcaacctcg ggcgagcagt cttccaggcc aaaaagaggc
     2461 ttcttgaacc tcttggtctg gttgaggaag cggctaagac ggctcctgga aagaagaggc
     2521 ctgtagagca gtctcctcag gaaccggact cctccgcggg tattggcaaa tcgggtgcac
     2581 agcccgctaa aaagagactc aatttcggtc agactggcga cacagagtca gtcccagacc
     2641 ctcaaccaat cggagaacct cccgcagccc cctcaggtgt gggatctctt acaatggctt
     2701 caggtggtgg cgcaccagtg gcagacaata acgaaggtgc cgatggagtg ggtagttcct
     2761 cgggaaattg gcattgcgat tcccaatggc tgggggacag agtcatcacc accagcaccc
     2821 gaacctgggc cctgcccacc tacaacaatc acctctacaa gcaaatctcc aacagcacat
     2881 ctggaggatc ttcaaatgac aacgcctact tcggctacag caccccctgg gggtattttg
     2941 acttcaacag attccactgc cacttctcac cacgtgactg gcagcgactc atcaacaaca
     3001 actggggatt ccggcctaag cgactcaact tcaagctctt caacattcag gtcaaagagg
     3061 ttacggacaa caatggagtc aagaccatcg ccaataacct taccagcacg gtccaggtct
     3121 tcacggactc agactatcag ctcccgtacg tgctcgggtc ggctcacgag ggctgcctcc
     3181 cgccgttccc agcggacgtt ttcatgattc ctcagtacgg gtatctgacg cttaatgatg
     3241 gaagccaggc cgtgggtcgt tcgtcctttt actgcctgga atatttcccg tcgcaaatgc
     3301 taagaacggg taacaacttc cagttcagct acgagtttga gaacgtacct ttccatagca
     3361 gctacgctca cagccaaagc ctggaccgac taatgaatcc actcatcgac caatacttgt
     3421 actatctctc tagaactatt aacggttctg gacagaatca acaaacgcta aaattcagtg
     3481 tggccggacc cagcaacatg gctgtccagg gaagaaacta catacctgga cccagctacc
     3541 gacaacaacg tgtctcaacc actgtgactc aaaacaacaa cagcgaattt gcttggcctg
     3601 gagcttcttc ttgggctctc aatggacgta atagcttgat gaatcctgga cctgctatgg
     3661 ccagccacaa agaaggagag gaccgtttct ttcctttgtc tggatcttta atttttggca
     3721 aacaaggtac cggtagagac aacgtggatg cggacaaagt catgataacc aacgaagaag
     3781 aaattaaaac tactaacccg gtagcaacgg agtcctatgg acaagtggcc acaaaccacc
     3841 agagtgccca acagaggcct ccgcgtgagc cggcacaggc gcagaccggt tgggttcaaa
     3901 accaaggaat acttccgggt atggtttggc aggacagaga tgtgtacctg caaggaccca
     3961 tttgggccaa aattcctcac acggacggca actttcaccc ttctccgctg atgggagggt
     4021 ttggaatgaa gcacccgcct cctcagatcc tcatcaaaaa cacacctgta cctgcggatc
     4081 ctccaacggc cttcaacaag gacaagctga actctttcat cacccagtat tctactggcc
     4141 aagtcagcgt ggagatcgag tgggagctgc agaaggaaaa cagcaagcgc tggaacccgg
     4201 agatccagta cacttccaac tattacaagt ctaataatgt tgaatttgct gttaatactg
     4261 aaggtgtata tagtgaaccc cgccccattg gcaccagata cctgactcgt aatctgtaag
     4321 tttaaacctg caggactaga ggtcctgtat tagaggtcac gtgagtgttt tgcgacattt
     4381 tgcgacacca tgtggtcacg ctgggtattt aagcccgagt gagcacgcag ggtctccatt
     4441 ttgaagcggg aggtttgaac gcgcagccgc caagccgaat tctgcagata tccatcacac
     4501 tggcggccgc tcgactagag cggccgccac cgcggtggag ctccagcttt tgttcccttt
     4561 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4621 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4681 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4741 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4801 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4861 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4921 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4981 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5041 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5101 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5161 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5221 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5281 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5341 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5401 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5461 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5521 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5581 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5641 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5701 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5761 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5821 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5881 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5941 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6001 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6061 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6121 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6181 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6241 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6301 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6361 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6421 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6481 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6541 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6601 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6661 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6721 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     6781 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     6841 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     6901 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     6961 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     7021 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     7081 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     7141 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     7201 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7261 gctattacgc cagctggcga aag
//
