LOCUS       sequence-260185-        2849 bp    DNA     circular SYN 14-APR-2025
DEFINITION  In vitro expression of H. influenzae protein D with C terminal 
            4xDQNAT glycosylation sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2849)
  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 2849)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-PD-4xDQNAT
FEATURES             Location/Qualifiers
     source          1..2849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..60
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        41..59
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             91..113
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1186..1203
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1258..1276)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1272..1319
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1337..2152)
                     /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     2060..2079
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2144..2163
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2274..2849,1..13)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2763..2782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagcctatgg aaaaacgcca gcaacgcgat cccgcgaaat taatacgact cactataggg
       61 agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg agatatacat
      121 atgaagtcag acaagatcat tatcgcgcat cgcggtgcct ccggctactt gccggagcat
      181 actttggaga gcaaagcctt agcatttgcc caacaagcgg attacttgga acaagacttg
      241 gccatgacta aggatgggcg tctggtggtg attcacgacc attttcttga tggtcttacg
      301 gatgtagcta agaagtttcc gcatcgccat cgtaaggacg ggcgctatta tgtaatcgat
      361 tttaccttga aggaaattca aagcttggag atgaccgaaa actttgagac gaaggatggg
      421 aagcaagcgc aagtctaccc gaatcgtttt cccttatgga aaagccattt tcgcatccat
      481 acttttgagg atgagattga gtttatccaa gggctggaaa agtctactgg caagaaagtt
      541 gggatctatc ctgagatcaa ggcaccgtgg tttcatcatc aaaacggtaa ggacatcgcc
      601 gctgagacct taaaagtact gaagaagtat ggctacgata aaaagactga catggtttac
      661 cttcaaacgt tcgattttaa tgagttgaaa cgtattaaaa cagaattatt gccccagatg
      721 gggatggatt tgaagttggt tcagttaatc gcctacaccg actggaagga aacgcaggag
      781 aaggatccca aaggatactg ggtgaattat aattatgatt ggatgttcaa gccaggagca
      841 atggctgaag tagtaaaata tgcggatggt gtaggccccg gctggtatat gcttgtgaac
      901 aaagaagaaa gcaagcctga taatatcgtc tatactccct tagtcaaaga actggcccaa
      961 tataacgtag aagttcaccc ttacaccgtc cgtaaggacg ccttaccgga gttttttact
     1021 gacgtcaatc aaatgtatga tgccttgctg aacaagagtg gagcgacagg cgtcttcacc
     1081 gactttccag acactctcga ggatcagaac gcgaccggcg gtgaccaaaa tgccacaggt
     1141 ggcgatcaaa acgccaccgg cggtgaccag aatgcgacag tcgaccatca ccatcatcac
     1201 cattaagtcg accggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     1261 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     1321 tgaaagccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     1381 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     1441 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     1501 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     1561 caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga
     1621 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     1681 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     1741 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     1801 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     1861 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     1921 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     1981 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     2041 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     2101 tggaatttaa tcgcggcttc gagcaagacg tttcccgttg aatatggctc ataacacccc
     2161 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     2221 gtgcaatgta acatcagaga ttttgagaca caacgtgaga tcaaaggatc ttcttgagat
     2281 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2341 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2401 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2461 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2521 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2581 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2641 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2701 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2761 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2821 cgatttttgt gatgctcgtc aggggggcg
//
