LOCUS       sequence-339786-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SunS expression in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6558)
  AUTHORS   Fujinami D, Garcia de Gonzalo CV, Biswas S, Hao Y, Wang H, Garg N, 
            Lukk T, Nair SK, van der Donk WA
  TITLE     Structural and mechanistic investigations of protein 
            S-glycosyltransferases.
  JOURNAL   Cell Chem Biol. 2021 Dec 16;28(12):1740-1749.e6. doi: 
            10.1016/j.chembiol.2021.06.009. Epub 2021 Jul 21.
  PUBMED    34283964
REFERENCE   2  (bases 1 to 6558)
  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"; doi: 
            "10.1016/j.chembiol.2021.06.009"; journalName: "Cell Chem Biol"; 
            date: "2021-12-16- 16"; volume: "28"; issue: "12"; pages: 
            "1740-1749.e6"
COMMENT     Sequence Label: pSunS
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1423..1440
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1493..1511)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1507..1554
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1591..2046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1678..1697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1888..1909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2138..2953)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2138..2953)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2861..2880
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2945..2964
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3075..3663
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3564..3583
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3817..3834
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3849..3988
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3914..3933)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4074..4096
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4090..4281)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5090..6172)
                     /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     6134..6153
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6173..6250)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6456..6475
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg aaactgagtg atatttattt ggaattaaag
      181 aaaggctatg ccgattcttt attgtattca gatttgtcat tgttggttaa tataatggaa
      241 tatgaaaaag atattgatgt gatgtcaatt caatctttgg ttgcaggtta tgaaaaatca
      301 gatactccta caataacatg cggtattata gtttataacg aaagcaagag aattaaaaag
      361 tgtttaaata gtgttaaaga tgattttaac gagattattg ttctagattc atactccact
      421 gatgataccg ttgatattat taaatgtgat tttcctgatg ttgaaattaa atatgaaaag
      481 tggaagaatg atttttccta tgctagaaat aaaattatag agtatgctac ttccgaatgg
      541 atttatttta ttgatgcaga taatttatac tctaaagaaa acaaagggaa aatagctaaa
      601 gtagctagag ttttagagtt tttttctatt gattgtgtag ttagtccata tatagaagaa
      661 tatactggac atctatattc tgatacacga agaatgtttc ggctcaatgg taaagttaaa
      721 tttcatggga aagtgcatga agaacctatg aattataatc atagtctacc ttttaatttc
      781 attgtgaacc ttaaggttta ccataatgga tataatcctt cagagaataa tataaaatca
      841 aaaacacgaa ggaatataaa tctcacagaa gaaatgttaa gattggagcc cgaaaaccca
      901 aaatggttat tctttttcgg cagagaacta catttacttg ataaagatga agaagcaatt
      961 gattatctga aaaaatcaat aaacaactat aaaaaattta atgatcaaag acattttata
     1021 gatgctttag tgctattatg tactttatta ttgcagagaa ataattatgt tgacttaact
     1081 ttatatttgg atatattgga aactgaatat ccaagatgtg ttgatgttga ttactttaga
     1141 tctgcaattt tgttagtaga tatgcaaaat aaacttactt ctttaagcaa tatgattgat
     1201 gaagctctta cagacgagag atacagtgct ataaatacaa caaaagatca ctttaaaaga
     1261 attttaataa gccttaatat tcaactcgaa aattgggaaa gagtaaaaga aatatcaggg
     1321 gaaattaaaa atgataatat gaaaaaagaa attaaacaat atcttgccaa ctcactccac
     1381 aatattgaac acgtcctgaa aggaattgaa gtatgactcg agcaccacca ccaccaccac
     1441 tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     1501 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1561 ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     1621 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1681 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1741 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1801 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1861 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1921 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1981 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     2041 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2101 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     2161 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     2221 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     2281 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     2341 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     2401 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     2461 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     2521 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     2581 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     2641 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     2701 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     2761 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     2821 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     2881 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     2941 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     3001 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3061 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3121 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3181 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3241 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3301 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3361 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3421 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3481 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3541 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3601 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3661 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3721 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3781 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3841 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     3901 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     3961 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     4021 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4081 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     4141 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     4201 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     4261 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     4321 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     4381 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     4441 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     4501 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     4561 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     4621 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     4681 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     4741 cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     4801 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     4861 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     4921 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     4981 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     5041 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     5101 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5161 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     5221 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     5281 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     5341 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
     5401 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     5461 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     5521 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     5581 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     5641 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     5701 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     5761 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     5821 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     5881 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     5941 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     6001 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     6061 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     6121 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     6181 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     6241 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     6301 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
     6361 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     6421 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     6481 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag
     6541 atctcgatcc cgcgaaat
//
