LOCUS       sequence-477386-        9333 bp    DNA     circular SYN 30-JUN-2025
DEFINITION  Express AAV2 capsid with ncAA mutant at site 454 of VP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9333)
  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 9333)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 30, 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-delVP2-VP2-454TAG
FEATURES             Location/Qualifiers
     source          1..9333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..22
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    235..282
                     /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             4231..4242
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4717..4733)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4726..4748)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     enhancer        4917..5296
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5297..5500
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5451..5471
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5545..5564
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5545..5563
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7221..7232
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(7634..7650)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7634..7650)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      7698..7729
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        7730..7832
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             7833..8648
                     /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(7906..7925)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      8672..8699
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(8711..9298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8791..8810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgtgtaaa acgacggcca gtttatctag tcagcttgat tctagctgat cgtggaccgg
       61 aaggtgagcc agtgagttga ttgcagtcca gttacgctgg agtctgaggc tcgtcctgaa
      121 tgatatgcga ccgccggagg gttgcgtttg agacgggcga cagatccagt cgcgctgctc
      181 tcgtcgatcc gctagggcgg ccgctctaga actagtggat cccccggaag atcagaagtt
      241 cctattccga agttcctatt ctctagaaag tataggaact tctgatctgc gcagccgcca
      301 tgccggggtt ttacgagatt gtgattaagg tccccagcga ccttgacgag catctgcccg
      361 gcatttctga cagctttgtg aactgggtgg ccgagaagga atgggagttg ccgccagatt
      421 ctgacatgga tctgaatctg attgagcagg cacccctgac cgtggccgag aagctgcagc
      481 gcgactttct gacggaatgg cgccgtgtga gtaaggcccc ggaggccctt ttctttgtgc
      541 aatttgagaa gggagagagc tacttccaca tgcacgtgct cgtggaaacc accggggtga
      601 aatccatggt tttgggacgt ttcctgagtc agattcgcga aaaactgatt cagagaattt
      661 accgcgggat cgagccgact ttgccaaact ggttcgcggt cacaaagacc agaaatggcg
      721 ccggaggcgg gaacaaggtg gtggatgagt gctacatccc caattacttg ctccccaaaa
      781 cccagcctga gctccagtgg gcgtggacta atatggaaca gtatttaagc gcctgtttga
      841 atctcacgga gcgtaaacgg ttggtggcgc agcatctgac gcacgtgtcg cagacgcagg
      901 agcagaacaa agagaatcag aatcccaatt ctgatgcgcc ggtgatcaga tcaaaaactt
      961 cagccaggta catggagctg gtcgggtggc tcgtggacaa ggggattacc tcggagaagc
     1021 agtggatcca ggaggaccag gcctcataca tctccttcaa tgcggcctcc aactcgcggt
     1081 cccaaatcaa ggctgccttg gacaatgcgg gaaagattat gagcctgact aaaaccgccc
     1141 ccgactacct ggtgggccag cagcccgtgg aggacatttc cagcaatcgg atttataaaa
     1201 ttttggaact aaacgggtac gatccccaat atgcggcttc cgtctttctg ggatgggcca
     1261 cgaaaaagtt cggcaagagg aacaccatct ggctgtttgg gcctgcaact accgggaaga
     1321 ccaacatcgc ggaggccata gcccacactg tgcccttcta cgggtgcgta aactggacca
     1381 atgagaactt tcccttcaac gactgtgtcg acaagatggt gatctggtgg gaggagggga
     1441 agatgaccgc caaggtcgtg gagtcggcca aagccattct cggaggaagc aaggtgcgcg
     1501 tggaccagaa atgcaagtcc tcggcccaga tagacccgac tcccgtgatc gtcacctcca
     1561 acaccaacat gtgcgccgtg attgacggga actcaacgac cttcgaacac cagcagccgt
     1621 tgcaagaccg gatgttcaaa tttgaactca cccgccgtct ggatcatgac tttgggaagg
     1681 tcaccaagca ggaagtcaaa gactttttcc ggtgggcaaa ggatcacgtg gttgaggtgg
     1741 agcatgaatt ctacgtcaaa aagggtggag ccaagaaaag acccgccccc agtgacgcag
     1801 atataagtga gcccaaacgg gtgcgcgagt cagttgcgca gccatcgacg tcagacgcgg
     1861 aagcttcgat caactacgca gacaggtacc aaaacaaatg ttctcgtcac gtgggcatga
     1921 atctgatgct gtttccctgc agacaatgcg agagaatgaa tcagaattca aatatctgct
     1981 tcactcacgg acagaaagac tgtttagagt gctttcccgt gtcagaatct caacccgttt
     2041 ctgtcgtcaa aaaggcgtat cagaaactgt gctacattca tcatatcatg ggaaaggtgc
     2101 cagacgcttg cactgcctgc gatctggtca atgtggattt ggatgactgc atctttgaac
     2161 aataaatgat ttaaatcagg tatggctgcc gatggttatc ttccagattg gctcgaggac
     2221 actctctctg aaggaataag acagtggtgg aagctcaaac ctggcccacc accaccaaag
     2281 cccgcagagc ggcataagga cgacagcagg ggtcttgtgc ttcctgggta caagtacctc
     2341 ggacccttca acggactcga caagggagag ccggtcaacg aggcagacgc cgcggccctc
     2401 gagcacgaca aagcctacga ccggcagctc gacagcggag acaacccgta cctcaagtac
     2461 aaccacgccg acgcggagtt tcaggagcgc cttaaagaag atacgtcttt tgggggcaac
     2521 ctcggacgag cagtcttcca ggcgaaaaag agggttcttg aacctctggg cctggttgag
     2581 gaacctgtta agaccgctcc gggaaaaaag aggccggtag agcactctcc tgtggagcca
     2641 gactcctcct cgggaaccgg aaaggcgggc cagcagcctg caagaaaaag attgaatttt
     2701 ggtcagactg gagacgcaga ctcagtacct gacccccagc ctctcggaca gccaccagca
     2761 gccccctctg gtctgggaac taatacgatg gctacaggca gtggcgcacc aatggcagac
     2821 aataacgagg gcgccgacgg agtgggtaat tcctcgggaa attggcattg cgattccaca
     2881 tggatgggcg acagagtcat caccaccagc acccgaacct gggccctgcc cacctacaac
     2941 aaccacctct acaaacaaat ttccagccaa tcaggagcct cgaacgacaa tcactacttt
     3001 ggctacagca ccccttgggg gtattttgac ttcaacagat tccactgcca cttttcacca
     3061 cgtgactggc aaagactcat caacaacaac tggggattcc gacccaagag actcaacttc
     3121 aagctcttta acattcaagt caaagaggtc acgcagaatg acggtacgac gacgattgcc
     3181 aataacctta ccagcacggt tcaggtgttt actgactcgg agtaccagct cccgtacgtc
     3241 ctcggctcgg cgcatcaagg atgcctcccg ccgttcccag cagacgtctt catggtgcca
     3301 cagtatggat acctcaccct gaacaacggg agtcaggcag taggacgctc ttcattttac
     3361 tgcctggagt actttccttc tcagatgctg cgtaccggaa acaactttac cttcagctac
     3421 acttttgagg acgttccttt ccacagcagc tacgctcaca gccagagtct ggaccgtctc
     3481 atgaatcctc tcatcgacca gtacctgtat tacttgagca gaacaaacac tccaagtgga
     3541 accaccacgc agtcaaggct tcagttttct caggccggag cgagtgacat tcgggaccag
     3601 tctaggaact ggcttcctgg accctgttac cgccagcagc gagtatcaaa gacatctgcg
     3661 gataacaaca acagtgaata ctcgtggact ggagctacca agtaccacct caatggcaga
     3721 gactctctgg tgaatccggg cccggccatg gcaagccaca aggacgatga agaaaagttt
     3781 tttcctcaga gcggggttct catctttggg aagcaaggct cagagaaaac aaatgtggac
     3841 attgaaaagg tcatgattac agacgaagag gaaatcagga caaccaatcc cgtggctacg
     3901 gagcagtatg gttctgtatc taccaacctc cagagaggca acagacaagc agctaccgca
     3961 gatgtcaaca cacaaggcgt tcttccaggc atggtctggc aggacagaga tgtgtacctt
     4021 caggggccca tctgggcaaa gattccacac acggacggac attttcaccc ctctcccctc
     4081 atgggtggat tcggacttaa acaccctcct ccacagattc tcatcaagaa caccccggta
     4141 cctgcgaatc cttcgaccac cttcagtgcg gcaaagtttg cttccttcat cacacagtac
     4201 tccacgggac aggtcagcgt ggagatcgag tgggagctgc agaaggaaaa cagcaaacgc
     4261 tggaatcccg aaattcagta cacttccaac tacaacaagt ctgttaatgt ggactttact
     4321 gtggacacta atggcgtgta ttcagagcct cgccccattg gcaccagata cctgactcgt
     4381 aatctgtaat tgcttgttaa tcaataaacc gtttaattcg tttcagttga actttggtct
     4441 ctgcgtattt ctttcttatc tagtttccat ggctacgtag ataagtagca tggcgggtta
     4501 atcattaact acagcccggg cgtttaaaca gcgggcggag gggtggagtc gtgacgtgaa
     4561 ttacgtcata gggttaggga ggtcctgtat tagaggtcac gtgagtgttt tgcgacattt
     4621 tgcgacacca tgtggtctcg ctgggggggg gggcccgagt gagcacgcag ggtctccatt
     4681 ttgaagcggg aggtttgaac gagcgctggc gcgctcactg gccgtcgttt tacaacgtcg
     4741 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     4801 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccatgcat cggccgcaaa
     4861 tacctgcagg atccgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     4921 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     4981 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5041 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5101 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5161 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5221 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     5281 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     5341 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     5401 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     5461 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     5521 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
     5581 gcatggctcc gggaaaaaag aggccggtag agcactctcc tgtggagcca gactcctcct
     5641 cgggaaccgg aaaggcgggc cagcagcctg caagaaaaag attgaatttt ggtcagactg
     5701 gagacgcaga ctcagtacct gacccccagc ctctcggaca gccaccagca gccccctctg
     5761 gtctcggaac taataccctc gctacaggca gtggcgcacc actcgcagac aataacgagg
     5821 gcgccgacgg agtgggtaat tcctcgggaa attggcattg cgattccaca tggctcggcg
     5881 acagagtcat caccaccagc acccgaacct gggccctgcc cacctacaac aaccacctct
     5941 acaaacaaat ttccagccaa tcaggagcct cgaacgacaa tcactacttt ggctacagca
     6001 ccccttgggg gtattttgac ttcaacagat tccactgcca cttttcacca cgtgactggc
     6061 aaagactcat caacaacaac tggggattcc gacccaagag actcaacttc aagctcttta
     6121 acattcaagt caaagaggtc acgcagaatg acggtacgac gacgattgcc aataacctta
     6181 ccagcacggt tcaggtgttt actgactcgg agtaccagct cccgtacgtc ctcggctcgg
     6241 cgcatcaagg atgcctcccg ccgttcccag cagacgtctt catggtgcca cagtatggat
     6301 acctcaccct gaacaacggg agtcaggcag taggacgctc ttcattttac tgcctggagt
     6361 actttccttc tcagatgctg cgtaccggaa acaactttac cttcagctac acttttgagg
     6421 acgttccttt ccacagcagc tacgctcaca gccagagtct ggaccgtctc atgaatcctc
     6481 tcatcgacca gtacctgtat tacttgagca gaacaaacac tccaagtgga tagaccacgc
     6541 agtcaaggct tcagttttct caggccggag cgagtgacat tcgggaccag tctaggaact
     6601 ggcttcctgg accctgttac cgccagcagc gagtatcaaa gacatctgcg gataacaaca
     6661 acagtgaata ctcgtggact ggagctacca agtaccacct caatggcaga gactctctgg
     6721 tgaatccggg cccggccatg gcaagccaca aggacgatga agaaaagttt tttcctcaga
     6781 gcggggttct catctttggg aagcaaggct cagagaaaac aaatgtggac attgaaaagg
     6841 tcatgattac agacgaagag gaaatcagga caaccaatcc cgtggctacg gagcagtatg
     6901 gttctgtatc taccaacctc cagagaggca acagacaagc agctaccgca gatgtcaaca
     6961 cacaaggcgt tcttccaggc atggtctggc aggacagaga tgtgtacctt caggggccca
     7021 tctgggcaaa gattccacac acggacggac attttcaccc ctctcccctc atgggtggat
     7081 tcggacttaa acaccctcct ccacagattc tcatcaagaa caccccggta cctgcgaatc
     7141 cttcgaccac cttcagtgcg gcaaagtttg cttccttcat cacacagtac tccacgggac
     7201 aggtcagcgt ggagatcgag tgggagctgc agaaggaaaa cagcaaacgc tggaatcccg
     7261 aaattcagta cacttccaac tacaacaagt ctgttaatgt ggactttact gtggacacta
     7321 atggcgtgta ttcagagcct cgccccattg gcaccagata cctgactcgt aatctgtaat
     7381 tgcttgttaa tcaataaacc gtttaattcg tttcagttga actttggtct ctgcgtattt
     7441 ctttcttatc tagtttccat ggctacgtag ataagtagca tggcgggtta atcattaact
     7501 acagccctag gggtgcgagc ggatcgagca gtgtcgatca ctactggacc gcgagctgtg
     7561 ctgcgacccg tgatcttacg gcattatacg tatgatcggt ccacgatcag ctagattatc
     7621 tagtcagctt gatgtcatag ctgtttcctg aggctcaata ctgaccattt aaatcatacc
     7681 tgacctccat agcagaaagt caaaagcctc cgaccggagg cttttgactt gatcggcacg
     7741 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     7801 tatcgagatt ttcaggagct aaggaagcta aaatgagcca tattcaacgg gaaacgtctt
     7861 gcttgaagcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     7921 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     7981 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     8041 tcaggctaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     8101 ctcctgatga tgcatggtta ctcaccactg cgatcccagg gaaaacagca ttccaggtat
     8161 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     8221 ggttgcattc gattcctgtt tgtaattgtc cttttaacgg cgatcgcgta tttcgtctcg
     8281 ctcaggcgca atcacgaatg aataacggtt tggttggtgc gagtgatttt gatgacgagc
     8341 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaactcttg ccattctcac
     8401 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     8461 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     8521 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     8581 aatatggtat tgataatcct gatatgaata aattgcagtt tcacttgatg ctcgatgagt
     8641 ttttctaatg aggacctaaa tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt
     8701 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa
     8761 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     8821 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     8881 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     8941 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     9001 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     9061 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     9121 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     9181 agccagttac ctcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     9241 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     9301 agatcctttg attttctacc gaagaaaggc cca
//
