LOCUS       sequence-260187-        3230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of tetanus toxin fragment C with C terminal 
            4xDQNAT glycosylation sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3230)
  AUTHORS   Stark JC, Jaroentomeechai T, Moeller TD, Hershewe JM, Warfel KF, 
            Moricz BS, Martini AM, Dubner RS, Hsu KJ, Stevenson TC, Jones BD, 
            DeLisa MP, Jewett MC
  TITLE     On-demand biomanufacturing of protective conjugate vaccines.
  JOURNAL   Sci Adv. 2021 Feb 3;7(6). pii: 7/6/eabe9444. doi: 
            10.1126/sciadv.abe9444. Print 2021 Feb.
  PUBMED    33536221
REFERENCE   2  (bases 1 to 3230)
  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: "Sci Adv."; 
            date: "2021-02-3"; volume: "7(6). pii: 7/6/eabe9444. doi"; pages: " 
            10.1126/sciadv.abe9444. Print 2021 Feb"
COMMENT     Sequence Label: pJL1-TTc-4xDQNAT
FEATURES             Location/Qualifiers
     source          1..3230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..58
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        39..57
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             89..111
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1565..1582
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1637..1655)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1651..1698
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1716..2531)
                     /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="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2439..2458
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2523..2542
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2653..3230,1..11)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3142..3161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctatggaa aaacgccagc aacgcgatcc cgcgaaatta atacgactca ctatagggag
       61 accacaacgg tttccctcta gaaataattt tgtttaactt taagaaggag atatacatat
      121 gaagaatctg gattgctggg tggacaatga agaagacatc gacgtgattc ttaaaaagag
      181 tacgatcctt aacttggata ttaacaatga cattatttca gacatctctg gtttcaatag
      241 ttcagtcatt acatatccgg atgcccaact ggttccagga atcaatggaa aggcgatcca
      301 ccttgtgaac aatgagtcta gcgaggtgat tgtccataag gctatggaca tcgaatacaa
      361 cgatatgttt aacaacttca cagtatcgtt ttggctgcgt gtaccaaaag tctcagcttc
      421 tcatttggaa caatacgata caaatgaata ctccatcatc agctctatga agaagtatag
      481 cctttcgatc ggcagtggtt ggtccgtgtc cttaaaaggt aataacttaa tctggacatt
      541 aaaagattca gccggcgaag tccgtcagat cactttccgc gacctttccg ataaatttaa
      601 cgcttactta gcaaacaaat gggtattcat taccatcaca aacgaccgtt tgagctctgc
      661 caatttgtac attaacggtg ttttaatggg tagcgcagag atcacgggcc ttggggctat
      721 ccgtgaggac aataatatca cacttaagct ggatcgctgc aataacaaca atcaatatgt
      781 gagcatcgac aaatttcgca ttttttgtaa ggctcttaac cccaaggaga tcgaaaagtt
      841 atatacgagt tacctgagca ttacctttct gcgcgacttt tgggggaatc cgttacgcta
      901 tgacacagaa tattatttaa tccctgtggc atactcttcg aaagacgtcc agctgaaaaa
      961 cattaccgac tatatgtact taacgaacgc tccaagctac acgaatggaa aattaaacat
     1021 ctattaccgt cgtttatatt ctggtctgaa atttatcatt aaacgctata cacccaataa
     1081 tgagattgat tcgttcgtcc gttccggtga cttcattaag ttgtatgttt cgtacaacaa
     1141 caacgaacat attgttgggt accccaaaga tggaaacgct ttcaataatt tggaccgcat
     1201 cttacgtgta ggctacaacg ctccagggat ccctctttac aaaaaaatgg aagcggtaaa
     1261 acttcgcgac ttaaaaactt attcagtcca gcttaagctt tatgacgata aagatgcctc
     1321 gcttggcctt gttgggactc ataacggcca aatcggtaat gaccccaacc gcgacatcct
     1381 tattgcatcc aattggtact ttaatcacct gaaggataag actctgactt gcgattggta
     1441 cttcgtaccg actgacgaag ggtggacgaa cgatctcgag gatcagaacg cgaccggcgg
     1501 tgaccaaaat gccacaggtg gcgatcaaaa cgccaccggc ggtgaccaga atgcgacagt
     1561 cgaccatcac catcatcacc attaagtcga ccggctgcta acaaagcccg aaaggaagct
     1621 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1681 gtcttgaggg gttttttgct gaaagccaat tctgattaga aaaactcatc gagcatcaaa
     1741 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1801 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1861 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1921 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     1981 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2041 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2101 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2161 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2221 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2281 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2341 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2401 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2461 tataaatcag catccatgtt ggaatttaat cgcggcttcg agcaagacgt ttcccgttga
     2521 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     2581 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtgagat
     2641 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2701 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2761 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2821 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2881 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2941 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3001 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3061 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3121 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3181 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
//
