LOCUS       sequence-477384-        7404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express AAV2-KO (R585,588A) capsid with ncAA mutant at site 454.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7404)
  AUTHORS   Erickson SB, Pham Q, Cao X, Glicksman J, Kelemen RE, Shahraeini SS, 
            Bodkin S, Kiyam Z, Chatterjee A
  TITLE     Precise Manipulation of the Site and Stoichiometry of Capsid 
            Modification Enables Optimization of Functional Adeno-Associated 
            Virus Conjugates.
  JOURNAL   Bioconjug Chem. 2024 Jan 17;35(1):64-71. doi: 
            10.1021/acs.bioconjchem.3c00411. Epub 2023 Dec 16.
  PUBMED    38103182
REFERENCE   2  (bases 1 to 7404)
  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"; doi: 
            "10.1021/acs.bioconjchem.3c00411"; journalName: "Bioconjug Chem"; 
            date: "2024-01-17- 17"; volume: "35"; issue: "1"; pages: "64-71"
COMMENT     Sequence Label: pIDTsmart-RC2-KO-454TAG
FEATURES             Location/Qualifiers
     source          1..7404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..25
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9..25
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    39..86
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4035..4046
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4521..4537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4530..4552)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     enhancer        4721..5100
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5101..5304
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5255..5275
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5349..5368
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5349..5367
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5509..5525)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5509..5525)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      5573..5604
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        5605..5707
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5708..6523
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5781..5800)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6547..6574
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(6586..7173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6666..6685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7214..7230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcggccgctc tagaactagt ggatcccccg gaagatcaga agttcctatt ccgaagttcc
       61 tattctctag aaagtatagg aacttctgat ctgcgcagcc gccatgccgg ggttttacga
      121 gattgtgatt aaggtcccca gcgaccttga cgagcatctg cccggcattt ctgacagctt
      181 tgtgaactgg gtggccgaga aggaatggga gttgccgcca gattctgaca tggatctgaa
      241 tctgattgag caggcacccc tgaccgtggc cgagaagctg cagcgcgact ttctgacgga
      301 atggcgccgt gtgagtaagg ccccggaggc ccttttcttt gtgcaatttg agaagggaga
      361 gagctacttc cacatgcacg tgctcgtgga aaccaccggg gtgaaatcca tggttttggg
      421 acgtttcctg agtcagattc gcgaaaaact gattcagaga atttaccgcg ggatcgagcc
      481 gactttgcca aactggttcg cggtcacaaa gaccagaaat ggcgccggag gcgggaacaa
      541 ggtggtggat gagtgctaca tccccaatta cttgctcccc aaaacccagc ctgagctcca
      601 gtgggcgtgg actaatatgg aacagtattt aagcgcctgt ttgaatctca cggagcgtaa
      661 acggttggtg gcgcagcatc tgacgcacgt gtcgcagacg caggagcaga acaaagagaa
      721 tcagaatccc aattctgatg cgccggtgat cagatcaaaa acttcagcca ggtacatgga
      781 gctggtcggg tggctcgtgg acaaggggat tacctcggag aagcagtgga tccaggagga
      841 ccaggcctca tacatctcct tcaatgcggc ctccaactcg cggtcccaaa tcaaggctgc
      901 cttggacaat gcgggaaaga ttatgagcct gactaaaacc gcccccgact acctggtggg
      961 ccagcagccc gtggaggaca tttccagcaa tcggatttat aaaattttgg aactaaacgg
     1021 gtacgatccc caatatgcgg cttccgtctt tctgggatgg gccacgaaaa agttcggcaa
     1081 gaggaacacc atctggctgt ttgggcctgc aactaccggg aagaccaaca tcgcggaggc
     1141 catagcccac actgtgccct tctacgggtg cgtaaactgg accaatgaga actttccctt
     1201 caacgactgt gtcgacaaga tggtgatctg gtgggaggag gggaagatga ccgccaaggt
     1261 cgtggagtcg gccaaagcca ttctcggagg aagcaaggtg cgcgtggacc agaaatgcaa
     1321 gtcctcggcc cagatagacc cgactcccgt gatcgtcacc tccaacacca acatgtgcgc
     1381 cgtgattgac gggaactcaa cgaccttcga acaccagcag ccgttgcaag accggatgtt
     1441 caaatttgaa ctcacccgcc gtctggatca tgactttggg aaggtcacca agcaggaagt
     1501 caaagacttt ttccggtggg caaaggatca cgtggttgag gtggagcatg aattctacgt
     1561 caaaaagggt ggagccaaga aaagacccgc ccccagtgac gcagatataa gtgagcccaa
     1621 acgggtgcgc gagtcagttg cgcagccatc gacgtcagac gcggaagctt cgatcaacta
     1681 cgcagacagg taccaaaaca aatgttctcg tcacgtgggc atgaatctga tgctgtttcc
     1741 ctgcagacaa tgcgagagaa tgaatcagaa ttcaaatatc tgcttcactc acggacagaa
     1801 agactgttta gagtgctttc ccgtgtcaga atctcaaccc gtttctgtcg tcaaaaaggc
     1861 gtatcagaaa ctgtgctaca ttcatcatat catgggaaag gtgccagacg cttgcactgc
     1921 ctgcgatctg gtcaatgtgg atttggatga ctgcatcttt gaacaataaa tgatttaaat
     1981 caggtatggc tgccgatggt tatcttccag attggctcga ggacactctc tctgaaggaa
     2041 taagacagtg gtggaagctc aaacctggcc caccaccacc aaagcccgca gagcggcata
     2101 aggacgacag caggggtctt gtgcttcctg ggtacaagta cctcggaccc ttcaacggac
     2161 tcgacaaggg agagccggtc aacgaggcag acgccgcggc cctcgagcac gacaaagcct
     2221 acgaccggca gctcgacagc ggagacaacc cgtacctcaa gtacaaccac gccgacgcgg
     2281 agtttcagga gcgccttaaa gaagatacgt cttttggggg caacctcgga cgagcagtct
     2341 tccaggcgaa aaagagggtt cttgaacctc tgggcctggt tgaggaacct gttaagacgg
     2401 ctccgggaaa aaagaggccg gtagagcact ctcctgtgga gccagactcc tcctcgggaa
     2461 ccggaaaggc gggccagcag cctgcaagaa aaagattgaa ttttggtcag actggagacg
     2521 cagactcagt acctgacccc cagcctctcg gacagccacc agcagccccc tctggtctgg
     2581 gaactaatac gatggctaca ggcagtggcg caccaatggc agacaataac gagggcgccg
     2641 acggagtggg taattcctcg ggaaattggc attgcgattc cacatggatg ggcgacagag
     2701 tcatcaccac cagcacccga acctgggccc tgcccaccta caacaaccac ctctacaaac
     2761 aaatttccag ccaatcagga gcctcgaacg acaatcacta ctttggctac agcacccctt
     2821 gggggtattt tgacttcaac agattccact gccacttttc accacgtgac tggcaaagac
     2881 tcatcaacaa caactgggga ttccgaccca agagactcaa cttcaagctc tttaacattc
     2941 aagtcaaaga ggtcacgcag aatgacggta cgacgacgat tgccaataac cttaccagca
     3001 cggttcaggt gtttactgac tcggagtacc agctcccgta cgtcctcggc tcggcgcatc
     3061 aaggatgcct cccgccgttc ccagcagacg tcttcatggt gccacagtat ggatacctca
     3121 ccctgaacaa cgggagtcag gcagtaggac gctcttcatt ttactgcctg gagtactttc
     3181 cttctcagat gctgcgtacc ggaaacaact ttaccttcag ctacactttt gaggacgttc
     3241 ctttccacag cagctacgct cacagccaga gtctggaccg tctcatgaat cctctcatcg
     3301 accagtacct gtattacttg agcagaacaa acactccaag tggatagacc acgcagtcaa
     3361 ggcttcagtt ttctcaggcc ggagcgagtg acattcggga ccagtctagg aactggcttc
     3421 ctggaccctg ttaccgccag cagcgagtat caaagacatc tgcggataac aacaacagtg
     3481 aatactcgtg gactggagct accaagtacc acctcaatgg cagagactct ctggtgaatc
     3541 cgggcccggc catggcaagc cacaaggacg atgaagaaaa gttttttcct cagagcgggg
     3601 ttctcatctt tgggaagcaa ggctcagaga aaacaaatgt ggacattgaa aaggtcatga
     3661 ttacagacga agaggaaatc aggacaacca atcccgtggc tacggagcag tatggttctg
     3721 tatctaccaa cctccaggcc ggcaacgccc aagcagctac cgcagatgtc aacacacaag
     3781 gcgttcttcc aggcatggtc tggcaggaca gagatgtgta ccttcagggg cccatctggg
     3841 caaagattcc acacacggac ggacattttc acccctctcc cctcatgggt ggattcggac
     3901 ttaaacaccc tcctccacag attctcatca agaacacccc ggtacctgcg aatccttcga
     3961 ccaccttcag tgcggcaaag tttgcttcct tcatcacaca gtactccacg ggacaggtca
     4021 gcgtggagat cgagtgggag ctgcagaagg aaaacagcaa acgctggaat cccgaaattc
     4081 agtacacttc caactacaac aagtctgtta atgtggactt tactgtggac actaatggcg
     4141 tgtattcaga gcctcgcccc attggcacca gatacctgac tcgtaatctg taattgcttg
     4201 ttaatcaata aaccgtttaa ttcgtttcag ttgaactttg gtctctgcgt atttctttct
     4261 tatctagttt ccatggctac gtagataagt agcatggcgg gttaatcatt aactacagcc
     4321 cgggcgttta aacagcgggc ggaggggtgg agtcgtgacg tgaattacgt catagggtta
     4381 gggaggtcct gtattagagg tcacgtgagt gttttgcgac attttgcgac accatgtggt
     4441 ctcgctgggg gggggggccc gagtgagcac gcagggtctc cattttgaag cgggaggttt
     4501 gaacgagcgc tggcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     4561 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4621 gaagaggccc gcaccgatcg cccttcccat gcatcggccg caaatacctg caggatccgt
     4681 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     4741 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     4801 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     4861 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     4921 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     4981 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     5041 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     5101 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     5161 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     5221 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     5281 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
     5341 atcgaaatta atacgactca ctatagggag acccaagctg gctaggggtg cgagcggatc
     5401 gagcagtgtc gatcactact ggaccgcgag ctgtgctgcg acccgtgatc ttacggcatt
     5461 atacgtatga tcggtccacg atcagctaga ttatctagtc agcttgatgt catagctgtt
     5521 tcctgaggct caatactgac catttaaatc atacctgacc tccatagcag aaagtcaaaa
     5581 gcctccgacc ggaggctttt gacttgatcg gcacgtaaga ggttccaact ttcaccataa
     5641 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     5701 agctaaaatg agccatattc aacgggaaac gtcttgcttg aagccgcgat taaattccaa
     5761 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     5821 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     5881 aggtagcgtt gccaatgatg ttacagatga gatggtcagg ctaaactggc tgacggaatt
     5941 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     6001 cactgcgatc ccagggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     6061 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     6121 ttgtcctttt aacggcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     6181 cggtttggtt ggtgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     6241 ctggaaagaa atgcataaac tcttgccatt ctcaccggat tcagtcgtca ctcatggtga
     6301 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     6361 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     6421 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     6481 gaataaattg cagtttcact tgatgctcga tgagtttttc taatgaggac ctaaatgtaa
     6541 tcacctggct caccttcggg tgggcctttc tgcgttgctg gcgtttttcc ataggctccg
     6601 cccccctgac gagcatcaca aaaatcgatg ctcaagtcag aggtggcgaa acccgacagg
     6661 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6721 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6781 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6841 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6901 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6961 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7021 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttacctcgg aaaaagagtt
     7081 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7141 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatttt ctaccgaaga
     7201 aaggcccacc cgtgtaaaac gacggccagt ttatctagtc agcttgattc tagctgatcg
     7261 tggaccggaa ggtgagccag tgagttgatt gcagtccagt tacgctggag tctgaggctc
     7321 gtcctgaatg atatgcgacc gccggagggt tgcgtttgag acgggcgaca gatccagtcg
     7381 cgctgctctc gtcgatccgc tagg
//
