LOCUS       sequence-393680-        2849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJL1 plasmid encoding H. influenzae protein D (PD) modified at the 
            C-terminus with 30 amino acids containing one optimal DQNAT sequon 
            followed by a 6x-His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2849)
  AUTHORS   Hershewe JM, Warfel KF, Iyer SM, Peruzzi JA, Sullivan CJ, Roth EW, 
            DeLisa MP, Kamat NP, Jewett MC
  TITLE     Improving cell-free glycoprotein synthesis by characterizing and 
            enriching native membrane vesicles.
  JOURNAL   Nat Commun. 2021 Apr 22;12(1):2363. doi: 10.1038/s41467-021-22329-3.
  PUBMED    33888690
REFERENCE   2  (bases 1 to 2849)
  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 
            Commun."; date: "2021-04-22"; volume: "12(1):2363. doi"; pages: " 
            10.1038/s41467-021-22329-3"
COMMENT     Sequence Label: pJL1-PD-DQNAT
FEATURES             Location/Qualifiers
     source          1..2849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     633..652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             683..705
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1703..1732
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1778..1795
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1850..1868)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1864..1911
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1929..2744)
                     /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     2652..2671
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2736..2755
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg atcccgcgaa attaatacga ctcactatag ggagaccaca
      661 acggtttccc tctagaaata attttgttta actttaagaa ggagatatac atatgaagtc
      721 agacaagatc attatcgcgc atcgcggtgc ctccggctac ttgccggagc atactttgga
      781 gagcaaagcc ttagcatttg cccaacaagc ggattacttg gaacaagact tggccatgac
      841 taaggatggg cgtctggtgg tgattcacga ccattttctt gatggtctta cggatgtagc
      901 taagaagttt ccgcatcgcc atcgtaagga cgggcgctat tatgtaatcg attttacctt
      961 gaaggaaatt caaagcttgg agatgaccga aaactttgag acgaaggatg ggaagcaagc
     1021 gcaagtctac ccgaatcgtt ttcccttatg gaaaagccat tttcgcatcc atacttttga
     1081 ggatgagatt gagtttatcc aagggctgga aaagtctact ggcaagaaag ttgggatcta
     1141 tcctgagatc aaggcaccgt ggtttcatca tcaaaacggt aaggacatcg ccgctgagac
     1201 cttaaaagta ctgaagaagt atggctacga taaaaagact gacatggttt accttcaaac
     1261 gttcgatttt aatgagttga aacgtattaa aacagaatta ttgccccaga tggggatgga
     1321 tttgaagttg gttcagttaa tcgcctacac cgactggaag gaaacgcagg agaaggatcc
     1381 caaaggatac tgggtgaatt ataattatga ttggatgttc aagccaggag caatggctga
     1441 agtagtaaaa tatgcggatg gtgtaggccc cggctggtat atgcttgtga acaaagaaga
     1501 aagcaagcct gataatatcg tctatactcc cttagtcaaa gaactggccc aatataacgt
     1561 agaagttcac ccttacaccg tccgtaagga cgccttaccg gagtttttta ctgacgtcaa
     1621 tcaaatgtat gatgccttgc tgaacaagag tggagcgaca ggcgtcttca ccgactttcc
     1681 agacactcta ggtgcggccg cagaacaaaa actcatctca gaagaggatc tgaatggggc
     1741 cgcactcgag ggtggcgatc agaacgcgac cggcggtcat caccatcatc accattaagt
     1801 cgaccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     1861 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagcc
     1921 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     1981 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     2041 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     2101 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     2161 gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca
     2221 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     2281 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     2341 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     2401 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     2461 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     2521 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     2581 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     2641 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     2701 aatcgcggct tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     2761 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     2821 taacatcaga gattttgaga cacaacgtg
//
