LOCUS       sequence-390797-        3050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of N-acylneuraminate cytodiylyltransfease 
            (NeuA), which conjugates sialic acid and CTP to make activated sugar
            donor, CMP-sialic acid (see CSTI.PJL1 and PdST6.PJL1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3050)
  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 3050)
  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: NeuA.pJL1
FEATURES             Location/Qualifiers
     source          1..3050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(293..340)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     336..354
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(409..432)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     RBS             1700..1722
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1753..1772)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1754..1772)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1800..2388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1880..1899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2499..2518)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(2510..3050,1..275)
                     /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     complement(2583..2602)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
       61 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
      121 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
      181 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
      241 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggctttc agcaaaaaac
      301 ccctcaagac ccgtttagag gccccaaggg gttatgctag ttattgctca gcggtggcag
      361 cagccaactc agcttccttt cgggctttgt tagcagccgg tcgacttatt tttcgaactg
      421 cggatggctc cacgcagatt taacgatctc ggcaatttcg ttttccagaa cggtataacc
      481 attgctgtta aagtgcaggc catcataggt atacattttg ttcaggttgc cataggaatc
      541 tttcagaaca tgatccaggc tgataaattt ggcgaaatcc acattttcac gcagatagct
      601 attcaggtca ttgatgatac gattatcacg atcaatacga ccaaaaacag gcggaacgct
      661 cagcaggtag attttgctgg ctgcattctt ttttttgatg tactggcagg tctttttcag
      721 ataccaaagg gtatcctctt ttttccagtc gctcacaaca atatcattgg tgccgaaaaa
      781 gataaacacg aattcgccaa aattaacaat gcgctctttc tcgatgatgt actcgtaata
      841 ttctttgctg ttgataccgg caatacccag gttattaact tcgatgtcgt tgatcttttt
      901 cacgtcccaa taatcaaaca ggctatgacc aatcagggta atatcgctca cgctatcaaa
      961 ttcgttgcgt ttttcgttga tgcgattatg gatgttctgg tacagaatct tctgacgatt
     1021 tttcttctgc tgaatggtaa tggccagttc aaaatccatg cgatcatcga tatccagtga
     1081 gctttcttta tccatgatgt aggccaggct ataacgaccg aaaaaatgtt tggtatgcag
     1141 ataatgctgt ttgttggcga taaaaatggc accattcgga tgatattcaa caatgctgtt
     1201 gcggttatat ttgctgtagt ccagatcaaa aaagctcagg gtgctataat catccagcgg
     1261 acgaataatc tggctcggtt tattgctacg ggtaacgcta acaacacact gatacttttc
     1321 cagggtctga tacagtttca cggcttcaat aatatgggtg ctatcacgaa acggactggt
     1381 cggctgcagc agtgcaaaat tttcataatc ggtatagatg ctcagggcat gtttgataaa
     1441 ttcaaagctg cttgctttat cggttgccag ttcttccgga cgcagcagaa aatctgcacc
     1501 atagctttcg gcaattgcac catactgttc gctgtcggtg gtaacaataa ctttttcaaa
     1561 catttcgctc tgcagtgctg cttcaatggt atatgccagc agcggtttat caatcagcat
     1621 cagtgcattt ttgttacgca gacctttgct accgctacgt gccggaataa ttgcgataat
     1681 tttggtgcgc atatgtatat ctccttctta aagttaaaca aaattatttc tagagggaaa
     1741 ccgttgtggt ctccctatag tgagtcgtat taatttcgcg ggatcgcgtt gctggcgttt
     1801 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1861 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1921 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1981 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2041 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2101 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2221 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2401 atctcacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2461 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2521 tcaacgggaa acgtcttgct cgaagccgcg attaaattcc aacatggatg ctgatttata
     2581 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     2641 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2701 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2761 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa
     2821 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2881 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2941 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3001 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
//
