LOCUS       sequence-260186-        2996 bp    DNA     circular SYN 15-APR-2025
DEFINITION  In vitro expression of N. meningitidis PorA with C terminal 4xDQNAT 
            glycosylation sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2996)
  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 2996)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-PorA-4xDQNAT
FEATURES             Location/Qualifiers
     source          1..2996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             949..966
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1021..1039)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1035..1082
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1100..1915)
                     /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     1823..1842
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1907..1926
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2037..2625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2526..2545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2653..2672
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2653..2671
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2703..2725
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 acccagtggg gtaaccgtga gtcttttatc gggttagcag gcgagttcgg aactttacgt
       61 gccgggcgcg tagcaaacca gtttgacgat gcctcccagg ccatcgaccc ctgggattct
      121 aataatgacg tagcaagcca actgggaatt tttaagcgcc acgatgatat gcctgtctcc
      181 gttcgttacg acagtccgga gtttagcggg ttctcgggat ctgtgcaatt cgttccgatc
      241 caaaacagta aatctgccta tacaccggca tactatacaa aaaacacaaa taataacttg
      301 accttagtac cagccgtggt tggaaagcca ggttctgatg tatattatgc gggactgaac
      361 tacaaaaacg gagggttcgc cggtaactat gcgtttaagt acgcccgcca cgctaacgtc
      421 ggccgtaacg cctttgaact gttcttgatt gggagcggat cggatcaagc gaaaggaaca
      481 gaccccctga aaaaccatca agtccaccgt ttgaccggag gatatgagga ggggggtttg
      541 aacttggcat tagccgcgca gttggactta tcggaaaacg gcgacaaaac taagaattcc
      601 actactgaaa tcgcagctac tgcgagctat cgttttggga atgcagtgcc tcgtatttca
      661 tacgctcacg gcttcgattt cattgagcgc ggcaaaaagg gagaaaatac gtcctacgac
      721 caaattatcg ccggtgtaga ttatgacttc tctaagcgta ctagcgcaat cgtgagcggc
      781 gcttggttga agcgcaatac gggtatcggg aattataccc aaatcaatgc cgcatctgtc
      841 ggcttgcgtc acaaatttct cgaggatcag aacgcgaccg gcggtgacca aaatgccaca
      901 ggtggcgatc aaaacgccac cggcggtgac cagaatgcga cagtcgacca tcaccatcat
      961 caccattaag tcgaccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1021 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1081 tgctgaaagc caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
     1141 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     1201 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     1261 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     1321 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc
     1381 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     1441 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     1501 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1561 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1621 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1681 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1741 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1801 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1861 tgttggaatt taatcgcggc ttcgagcaag acgtttcccg ttgaatatgg ctcataacac
     1921 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     1981 cttgtgcaat gtaacatcag agattttgag acacaacgtg agatcaaagg atcttcttga
     2041 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2101 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2161 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2221 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2281 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2341 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2401 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2461 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2521 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2581 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2641 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
     2701 attttgttta actttaagaa ggagatatac atatggacgt cagtttgtat ggagagatta
     2761 aggcaggggt tgaggggcgc aattaccaac ttcaacttac ggaagcgcag gcagctaatg
     2821 gtggggcgag tggccaggtc aaagtcacga aggttactaa ggcaaagtcc cgcattcgta
     2881 cgaagatttc cgactttggc tcatttattg gtttcaaggg ttccgaggac ttaggcgatg
     2941 gcttgaaagc agtttggcag cttgagcaag atgtgagtgt ggcgggaggg ggcgct
//
