LOCUS       sequence-393684-        2883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJL1 plasmid encoding wildtype AcrA from C. jejuni containing 2 
            native internal glycosylation sites recognized by PglB and a 
            C-terminal 6x-His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2883)
  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 2883)
  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-AcrA
FEATURES             Location/Qualifiers
     source          1..2883
                     /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             1751..1768
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1782..1799
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1824..1841
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1884..1902)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1898..1945
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1963..2778)
                     /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     2686..2705
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2770..2789
                     /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 atatgaaaga
      721 agaagcacca aaaatacaaa tgccgcctca acctgtaaca accatgagtg ctaaatctga
      781 agatttacca cttagtttta cttaccctgc taaacttgtc agtgattatg atgtcattat
      841 aaaacctcaa gttagcggcg taatagtaaa taaacttttt aaagctggag ataaggtaaa
      901 aaaaggacaa acattattta ttatagaaca agataaattt aaagctagtg ttgattcagc
      961 ttacggacag gctttaatgg ctaaggcaac tttcgaaaat gcaagcaagg attttaatcg
     1021 ttctaaagct ctttttagca aaagtgcaat ctctcaaaaa gaatacgact cttctcttgc
     1081 tacatttaac aattcaaaag ctagtctagc aagtgctaga gcacagcttg caaatgcaag
     1141 aattgatcta gatcataccg agataaaagc tccttttgat ggtactatag gagatgcttt
     1201 agttaatata ggagattatg taagtgcttc aacaactgaa ctagttagag ttacagattt
     1261 aaatcctatt tacgcagatt tctttatttc agatacagat aaactaaatt tagtccgcaa
     1321 tactcaaagt ggaaaatggg atttagacag cattcatgca aatttaaatc ttaatggaga
     1381 aaccgttcaa ggcaaacttt attttattga ttcggttata gatgctaata gtggaacagt
     1441 aaaagccaaa gccgtatttg ataacaataa ctcaacactt ttaccgggtg cttttgcaac
     1501 aattacctca gaaggtttta tacaaaaaaa tggctttaaa gtgcctcaaa taggtgttaa
     1561 acaagatcaa aatgatgttt atgttcttct cgttaaaaat ggaaaagtag aaaaatcttc
     1621 tgtacatata agctaccaaa acaatgaata cgccattatt gacaaaggat tgcaaaatgg
     1681 cgataaaatc attttagata actctaaaaa aattcaagtt ggtagcgaag ttaaagaaat
     1741 tggagcacaa caccatcacc atcaccatta agaccggcgg tcatcaccat catcaccatt
     1801 aaaagcttgc ggccgcactc gagcaccacc accaccacca ctgagatccg gctgctaaca
     1861 aagccgtcga ccggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     1921 taaacgggtc ttgaggggtt ttttgctgaa agccaattct gattagaaaa actcatcgag
     1981 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     2041 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     2101 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     2161 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     2221 caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     2281 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     2341 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     2401 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     2461 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     2521 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     2581 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     2641 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     2701 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcttcgagc aagacgtttc
     2761 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     2821 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac
     2881 gtg
//
