LOCUS       sequence-260189-        3245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of tetanus toxin light chain with C terminal 
            4xDQNAT glycosylation sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3245)
  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 3245)
  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-TTlight-4xDQNAT
FEATURES             Location/Qualifiers
     source          1..3245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..49
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        30..48
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             80..102
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1571..1588
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1643..1661)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1657..1704
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1722..2537)
                     /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     2445..2464
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2529..2548
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2659..3245,1..2)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3148..3167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaacgccag caacgcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg
       61 gtttccctct agaaataatt ttgtttaact ttaagaagga gatatacata tgcctatcac
      121 catcaataac ttccgctact cagatccagt gaacaacgac actattatta tgatggaacc
      181 cccctattgc aagggattag atatctatta taaagccttc aagattaccg atcgtatctg
      241 gattgtcccc gagcgctacg aattcgggac caaacccgaa gattttaatc caccaagcag
      301 cttgatcgag ggtgcttcgg agtactatga tcccaactac ctgcgtaccg attcggataa
      361 agaccgtttt ctgcagacga tggtaaaact tttcaatcgt attaagaata atgtcgcggg
      421 cgaagcctta ttggataaga ttatcaatgc gattccttat cttggcaaca gttactcgtt
      481 actggacaag tttgatacga attcaaatag cgtgagtttt aacctgttgg aacaggaccc
      541 ttcaggcgcg accactaaaa gcgcaatgtt aaccaactta atcatttttg gacccggtcc
      601 cgtattgaac aaaaatgagg ttcgcgggat cgtactgcgt gtagacaata agaattactt
      661 tccgtgccgc gatggctttg gctcgattat gcaaatggcg ttttgcccag agtacgtgcc
      721 aacattcgac aatgtcatcg aaaatatcac ctctctgacc atcggaaaga gtaaatattt
      781 ccaagatcct gctttgttgc tgatgcatgc cctgatccat gtgttacatg gtttatacgg
      841 tatgcaagtt tcttcgcacg aaattattcc ctcgaaacag gaaatttaca tgcaacatac
      901 atacccgatt agtgcggagg agctgttcac ctttggcggg caggacgcga atttaatctc
      961 tattgatatc aagaacgact tatatgaaaa gacattgaac gactacaaag caatcgcaaa
     1021 caaactgagt caggtcacct catgcaatga cccgaacatt gatattgaca gttataagca
     1081 gatctaccaa caaaaatacc aatttgataa ggattctaat ggtcaatata ttgtgaacga
     1141 ggataagttt cagattcttt ataatagcat catgtatggt tttactgaga ttgagctggg
     1201 gaaaaaattc aacattaaaa cccgtctttc gtatttttcg atgaaccacg atcccgttaa
     1261 aattccgaac ttactggatg acactatcta taacgacacc gaaggattta acattgagag
     1321 caaggatctg aagtctgaat acaagggcca aaacatgcgc gtaaatacga atgccttccg
     1381 caatgtcgat ggatcgggac tggtgagcaa attgatcggc ttatgtaaga agattatccc
     1441 gcctacaaac atccgtgaga acttatacaa ccgcactgcc ctcgaggatc agaacgcgac
     1501 cggcggtgac caaaatgcca caggtggcga tcaaaacgcc accggcggtg accagaatgc
     1561 gacagtcgac catcaccatc atcaccatta agtcgaccgg ctgctaacaa agcccgaaag
     1621 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     1681 aaacgggtct tgaggggttt tttgctgaaa gccaattctg attagaaaaa ctcatcgagc
     1741 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     1801 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     1861 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     1921 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     1981 aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     2041 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     2101 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     2161 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     2221 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     2281 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     2341 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     2401 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     2461 tacccatata aatcagcatc catgttggaa tttaatcgcg gcttcgagca agacgtttcc
     2521 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     2581 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg
     2641 tgagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2701 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2761 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2821 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2881 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2941 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3001 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3061 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3121 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3181 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3241 atgga
//
