LOCUS       sequence-205287-        6334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses truncated PglF from C. jejuni with N-terminal GST tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6334)
  AUTHORS   Zamora CY, Schocker NS, Chang MM, Imperiali B
  TITLE     Chemoenzymatic Synthesis and Applications of Prokaryote-Specific 
            UDP-Sugars.
  JOURNAL   Methods Enzymol. 2017;597:145-186. doi: 10.1016/bs.mie.2017.06.003. 
            Epub 2017 Jul 5.
  PUBMED    28935101
REFERENCE   2  (bases 1 to 6334)
  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: "Methods 
            Enzymol."; date: "2017"; volume: "597:145-186. doi"; pages: " 
            10.1016/bs.mie.2017.06.003. Epub 2017 Jul 5"
COMMENT     Sequence Label: GST-PglF130
FEATURES             Location/Qualifiers
     source          1..6334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2202..2224)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2369..2387)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2455..2559
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2560..2628,2629..3420)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2778..2797)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3591..4179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4080..4099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4333..4350
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4423..4500
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4501..5583
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4520..4539)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5632..5637,5638..5655,5656..5662)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5670..5686
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5694..5710
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5694..5710
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5706..6233
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5726..5743)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5726..5742)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5735..5757)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tccatgcttg tggattttaa accttctaga
      781 atgaaagaag aagaaactcc ttgtattgta gtaggggcaa cttctaaggc tttgcatttg
      841 ttaaaaggtg caaaagaagg ttctttaggg ctttttcctg taggcgtagt tgatgcgaga
      901 aaagagctta tagggactta ttgtgataaa tttattgtag aagaaaaaga aaaaataaaa
      961 tcttatgtag aacaaggggt aaaaactgcc attattgctt taagacttga acaagaagag
     1021 cttaaaaaac tttttgaaga acttgtagct tatggtattt gcgatgtaaa aatattttct
     1081 tttacaagaa acgaagcaag agatatcagt atagaagact tgcttgctag aaaaccaaaa
     1141 gatttagatg atagtgctgt ggcggctttt ttaaaagata aggtagtttt ggtaagtgga
     1201 gcaggtggaa ctataggcag tgaactttgt aagcaatgta ttaaatttgg tgctaagcat
     1261 cttatcatgg ttgatcatag tgagtataat ctttataaga tcaatgatga tttaaattta
     1321 tataaagaaa aaattactcc tattttactg agtattttag ataagcaaag tttagatgag
     1381 gtattaaaaa cttataaacc cgagcttatt ttacatgcag ccgcttataa acatgtgcct
     1441 ctttgcgaac aaaatccaca ttcagcagta atcaataata ttttaggaac taaaatttta
     1501 tgcgacagtg ctaaagaaaa caaagtagct aaatttgtga tgataagtac agataaagca
     1561 gtacgaccaa caaatattat gggttgcact aagagagttt gcgagcttta tactttaagt
     1621 atgagtgatg aaaattttga agttgcttgt gtgcgttttg gtaatgtttt aggttctagt
     1681 ggtagtgtga taccgaaatt taaagcacaa attgccaata atgagccttt aactttaacg
     1741 caccctgata tagtgcgtta ttttatgctt gtggctgagg cagtgcaact tgttttacaa
     1801 gctggagcta tcgcaaaagg gggagaactt tttgttttgg atatgggtaa gcctgtgaaa
     1861 atcatagatt tagctaaaaa aatgctttta ctttctaatc gcaatgattt agaaattaaa
     1921 atcacaggct taagaaaagg tgagaagctt tatgaagagc ttttgattga tgaaaatgat
     1981 gctaaaacac aatatgagag tatttttgta gcaaagaatg agaaggttga ccttgattgg
     2041 cttaataaag agatagaaaa tttacaaata tgtgaagata tttcagaggc tttattaaag
     2101 attgtacctg aatttaaaca caataaagaa ggtgtactcg agcggccgca tcgtgactga
     2161 ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2221 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2281 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     2341 ggagtgtata attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2401 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2461 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2521 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2581 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2641 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2701 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2761 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     2821 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2881 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2941 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3001 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3061 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     3121 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3181 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3241 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3301 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3361 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3421 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3481 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3541 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3601 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3661 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3721 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3781 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3841 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3901 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3961 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4021 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4081 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4141 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4201 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4261 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4321 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     4381 ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggcgc
     4441 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     4501 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     4561 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     4621 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     4681 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     4741 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     4801 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     4861 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     4921 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     4981 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     5041 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     5101 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     5161 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     5221 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     5281 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     5341 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     5401 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     5461 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     5521 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5581 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg ctttacactt
     5641 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5701 cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     5761 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     5821 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     5881 ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg gagtgcgatc
     5941 ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt tacgatgcgc
     6001 ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt cccacggaga
     6061 atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc
     6121 agacgcgaat tatttttgat ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg
     6181 cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg
     6241 cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc
     6301 ataacggttc tggcaaatat tctgaaatga gctg
//
