LOCUS       sequence-386238-        7283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV packaging plasmid, expressing Rep2 and AAV-MG1.1 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/MG1.1
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      91..546
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(178..197)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     388..409
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        572..676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(677..745,746..1537)
                     /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     complement(895..914)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1708..2296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2197..2216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2450..2467
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2584..2605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2620..2625,2626..2643,2644..2650)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2658..2674
                     /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)."
     primer_bind     2663..2685
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2682..2698
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2682..2698
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2717..2737
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2719..2737
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(3121..3132)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(7142..7158)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7143..7159)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(7187..7206)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7188..7206)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7216..7233)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7216..7232)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7225..7247)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
       61 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
      121 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
      181 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
      241 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
      301 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
      361 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
      421 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
      481 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
      541 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      601 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      661 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      721 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      781 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      841 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      901 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      961 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1021 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1081 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1141 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1201 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     1261 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1321 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1381 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1441 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1501 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1561 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1621 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1681 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1741 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1801 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1861 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1921 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1981 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2041 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2101 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2161 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2221 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2281 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2341 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2401 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2461 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2521 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2581 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     2641 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     2701 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctccaccg
     2761 cggtggcggc cgctctagtc gagcggccgc cagtgtgatg gatatctgca gaattcggct
     2821 tggcggctgc gcgttcaaac ctcccgcttc aaaatggaga ccctgcgtgc tcactcgggc
     2881 ttaaataccc agcgtgacca catggtgtcg caaaatgtcg caaaacactc acgtgacctc
     2941 taatacagga cctctagtcc tgcaggttta aacttacaga ttacgagtca ggtatctggt
     3001 gccaatgggg cggggttcac tatatacacc ttcagtatta acagcaaatt caacattatt
     3061 agacttgtaa tagttggaag tgtactggat ctccgggttc cagcgcttgc tgttttcctt
     3121 ctgcagctcc cactcgatct ccacgctgac ttggccagta gaatactggg tgatgaaaga
     3181 gttcagcttg tccttgttga aggccgttgg aggatccgca ggtacaggtg tgtttttgat
     3241 gaggatctga ggaggcgggt gcttcattcc aaaccctccc atcagcggag aagggtgaaa
     3301 gttgccgtcc gtgtgaggaa ttttggccca aatgggtcct tgcaggtaca catctctgtc
     3361 ctgccaaacc atacccggaa gtattccttg gttttgaacc caaccggtct gcgcctgtgc
     3421 agacgtagtc ttcggcggag tcatcagact ctggtggttt gtggccactt gtccatagga
     3481 ctccgttgct accgggttag tagttttaat ttcttcttcg ttggttatca tgactttgtc
     3541 cgcatccacg ttgtctctac cggtaccttg tttgccaaaa attaaagatc cagacaaagg
     3601 aaagaaacgg tcctctcctt ctttgtggct ggccatagca ggtccaggat tcatcaagct
     3661 attacgtcca ttgagagccc aagaagaagc tccaggccaa gcaaattcgc tgttgttgtt
     3721 ttgagtcaca gtggttgaga cacgttgttg tcggtagctg ggtccaggta tgtagtttct
     3781 tccctggaca gccatgttgc tgggtccggc cacactgaat tttagcgttt gttgattctg
     3841 tccagaaccg ttaatagttc tagagagata gtacaagtat tggtcgatga gtggattcat
     3901 tagtcggtcc aggctttggc tgtgagcgta gctgctatgg aaaggtacgt tctcaaactc
     3961 gtagctgaac tggaagttgt tacccgttct tagcatttgc gacgggaaat attccaggca
     4021 gtaaaaggac gaacgaccca cggcctggct tccatcatta agcgtcagat acccgtactg
     4081 aggaatcatg aaaacgtccg ctgggaacgg cgggaggcag ccctcgtgag ccgacccgag
     4141 cacgtacggg agctgatagt ctgagtccgt gaagacctgg accgtgctgg taaggttatt
     4201 ggcgatggtc ttgactccat tgttgtccgt aacctctttg acctgaatgt tgaagagctt
     4261 gaagttgagt cgcttaggcc ggaatcccca gttgttgttg atgagtcgct gccagtcacg
     4321 tggtgagaag tggcagtgga atctgttgaa gtcaaaatac ccccaggggg tgctgtagcc
     4381 gaagtaggcg ttgtcatttg aagatcctcc agatgtgctg ttggagattt gcttgtagag
     4441 gtgattgttg taggtgggca gggcccaggt tcgggtgctg gtggtgatga ctctgtcccc
     4501 cagccattgg gaatcgcaat gccaatttcc cgaggaacta cccactccat cggcaccttc
     4561 gttattgtct gccactggtg cgccaccacc tgaagccatt gtaagagatc ccacacctga
     4621 gggggctgcg ggaggttctc cgattggttg agggtctggg actgactctg tgtcgccagt
     4681 ctgaccgaaa ttgagtctct ttttagcggg ctgtgcaccc gatttgccaa tacccgcgga
     4741 ggagtccggt tcctgaggag actgctctac aggcctcttc tttccaggag ccgtcttagc
     4801 cgcttcctca accagaccaa gaggttcaag aagcctcttt ttggcctgga agactgctcg
     4861 cccgaggttg cccccaaaag acgtatcttc tttgagccgc tcctggaact cggcgtcggc
     4921 gtggttgtac ttgaggtacg ggttgtctcc ggccttgagc tgctggtcgt aggccttgtc
     4981 gtgctcgagg gccgccgcgt ctgctgcgtt gaccggctcc cccttgtcga gtccgttgcc
     5041 gggtccaagg tatttgtaac ccggaagcac aagacctcga gcgttgtctt gatgttgttg
     5101 atttgccttg ggttgagggg ctccaggttt caaagcccac cactcgcgaa ttccttcact
     5161 aaggttgtcc tcgagccaat ctggaagata accatcggca gccatacctg atttaaatca
     5221 tttattgttc aaagatgcag tcatccaaat ccacattgac cagatcgcag gcagtgcaag
     5281 cgtctggcac ctttcccatg atatgatgaa tgtagcacag tttctgatac gcctttttga
     5341 cgacagaaac gggttgagat tctgacacgg gaaagcactc taaacagtct ttctgtccgt
     5401 gagtgaagca gatatttgaa ttctgattca ttctctcgca ttgtctgcag ggaaacagca
     5461 tcagattcat gcccacgtga cgagaacatt tgttttggta cctgtctgcg tagttgatcg
     5521 aagcttccgc gtctgacgtc gatggctgcg caactgactc gcgcacccgt ttgggctcac
     5581 ttatatctgc gtcactgggg gcgggtcttt tcttggctcc accctttttg acgtagaatt
     5641 catgctccac ctcaaccacg tgatcctttg cccaccggaa aaagtctttg acttcctgct
     5701 tggtgacctt cccaaagtca tgatccagac ggcgggtgag ttcaaatttg aacatccggt
     5761 cttgcaacgg ctgctggtgt tcgaaggtcg ttgagttccc gtcaatcacg gcgcacatgt
     5821 tggtgttgga ggtgacgatc acgggagtcg ggtctatctg ggccgaggac ttgcatttct
     5881 ggtccacgcg caccttgctt cctccgagaa tggctttggc cgactccacg accttggcgg
     5941 tcatcttccc ctcctcccac cagatcacca tcttgtcgac acagtcgttg aagggaaagt
     6001 tctcattggt ccagtttacg cacccgtaga agggcacagt gtgggctatg gcctccgcga
     6061 tgttggtctt cccggtagtt gcaggcccaa acagccagat ggtgttcctc ttgccgaact
     6121 ttttcgtggc ccatcccaga aagacggaag ccgcatattg gggatcgtac ccgtttagtt
     6181 ccaaaatttt ataaatccga ttgctggaaa tgtcctccac gggctgctgg cccaccaggt
     6241 agtcgggggc ggttttagtc aggctcataa tctttcccgc attgtccaag gcagccttga
     6301 tttgggaccg cgagttggag gccgcattga aggagatgta tgaggcctgg tcctcctgga
     6361 tccactgctt ctccgaggta atccccttgt ccacgagcca cccgaccagc tccatgtacc
     6421 tggctgaagt ttttgatctg atcaccggcg catcagaatt gggattctga ttctctttgt
     6481 tctgctcctg cgtctgcgac acgtgcgtca gatgctgcgc caccaaccgt ttacgctccg
     6541 tgagattcaa acaggcgctt aaatactgtt ccatattagt ccacgcccac tggagctcag
     6601 gctgggtttt ggggagcaag taattgggga tgtagcactc atccaccacc ttgttcccgc
     6661 ctccggcgcc atttctggtc tttgtgaccg cgaaccagtt tggcaaagtc ggctcgatcc
     6721 cgcggtaaat tctctgaatc agtttttcgc gaatctgact caggaaacgt cccaaaacca
     6781 tggatttcac cccggtggtt tccacgagca cgtgcatgtg gaagtagctc tctcccttct
     6841 caaattgcac aaagaaaaga gcctccgggg ccttactcac acggcgccat tccgtcagaa
     6901 agtcgcgctg cagcttctcg gccacggtca ggggtgcctg ctcaatcaga ttcagatcca
     6961 tgtcagaatc tggcggcaac tcccattcct tctcggccac ccagttcaca aagctgtcag
     7021 aaatgccggg cagatgctcg tcaaggtcgc tggggacctt aatcacaatc tcgtaaaacc
     7081 ccggcatggc ggctgcgcgt tcaaacctcc cgcttcaaaa tggagaccct gcgagctccc
     7141 ccgataccgt cgacctcgat cgaggggggg cccggtaccc aattcgccct atagtgagtc
     7201 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     7261 tacccaactt aatcgccttg cag
//
