LOCUS       sequence-390798-        2681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of glycosyltransferase from Campylobacter jejuni
            for elaboration of sialic acid onto galactose in alpha-2,3 linkage 
            using CMP-sialic acid as substrate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2681)
  AUTHORS   Thames AH, Moons SJ, Wong DA, Boltje TJ, Bochner BS, Jewett MC
  TITLE     GlycoCAP: A Cell-Free, Bacterial Glycosylation Platform for Building
            Clickable Azido-Sialoglycoproteins.
  JOURNAL   ACS Synth Biol. 2023 Apr 11. doi: 10.1021/acssynbio.3c00017.
  PUBMED    37040463
REFERENCE   2  (bases 1 to 2681)
  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: "ACS Synth 
            Biol. 2023 Apr 11. doi: 10.1021/acssynbio.3c00017."
COMMENT     Sequence Label: CSTI.pJL1
FEATURES             Location/Qualifiers
     source          1..2681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      107..154
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(172..987)
                     /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     895..914
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     979..998
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1109..1697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1598..1617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1725..1744
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1725..1743
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1775..1797
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1820..1843
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
ORIGIN
        1 agctgaaagc cgataacaaa attatgctga acttttatta agtcgaccgg ctgctaacaa
       61 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
      121 tggggcctct aaacgggtct tgaggggttt tttgctgaaa gccaattctg attagaaaaa
      181 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
      241 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
      301 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
      361 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
      421 tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
      481 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
      541 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
      601 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
      661 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
      721 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
      781 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
      841 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
      901 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcttcgagca
      961 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     1021 cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg
     1081 agacacaacg tgagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1141 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1201 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1261 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1321 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1381 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1441 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1501 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1561 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1621 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1681 ggcggagcct atggaaaaac gccagcaacg cgatcccgcg aaattaatac gactcactat
     1741 agggagacca caacggtttc cctctagaaa taattttgtt taactttaag aaggagatat
     1801 acatatggag aaaaaaatct ggagccatcc gcagttcgaa aaaggcggat ccggaggcag
     1861 ccacatgacc cgtacccgta tggaaaatga actgattgtg agcaaaaaca tgcagaacat
     1921 tatcattgca ggtaatggtc cgagcctgaa aaacattaac tataaacgtc tgcctcgcga
     1981 gtatgatgtt tttcgttgta accagttcta tttcgaggat aaatactatc tgggcaaaaa
     2041 aatcaaagcc gtgtttttca atccgggtgt ttttctgcag cagtatcata ccgcaaaaca
     2101 gctgattctg aaaaacgagt acgagatcaa aaacatcttt tgcagcacct ttaacctgcc
     2161 gtttattgaa agcaacgatt tcctgcacca gttttacaac ttttttccgg atgcaaaact
     2221 gggctatgaa gtgattgaaa acctgaaaga gttctacgcc tatatcaaat acaacgagat
     2281 ctatttcaac aaacgcatta ccagcggtgt ttatatgtgt gcaattgcca ttgccctggg
     2341 ctataaaacc atttatctgt gcggtatcga tttctatgaa ggcgacgtta tttatccgtt
     2401 tgaagcaatg agcaccaaca ttaaaacaat cttccctggc atcaaagact tcaaaccgag
     2461 caattgtcac agcaaagaat atgatattga ggccctgaaa ctgctgaaaa gcatctataa
     2521 agtgaacatc tatgccctgt gtgatgatag cattctggca aatcattttc cgctgagcat
     2581 taacatcaac aacaacttta ccctggaaaa caaacacaac aacagcatca atgatatcct
     2641 gctgaccgat aatacaccgg gtgttagctt ttacaaaaat c
//
