LOCUS       sequence-434038-        5805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heterologous expression of SSAT1 for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5805)
  AUTHORS   Mayers JR, Varon J, Zhou RR, Daniel-Ivad M, Beaulieu C, Bholse A, 
            Glasser NR, Lichtenauer FM, Ng J, Vera MP, Huttenhower C, Perrella 
            MA, Clish CB, Zhao SD, Baron RM, Balskus EP
  TITLE     A metabolomics pipeline highlights microbial metabolism in 
            bloodstream infections.
  JOURNAL   Cell. 2024 Jun 11:S0092-8674(24)00579-8. doi: 
            10.1016/j.cell.2024.05.035.
  PUBMED    38885650
REFERENCE   2  (bases 1 to 5805)
  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: "Cell. 2024 
            Jun 11:S0092-8674(24)00579-8. doi: 10.1016/j.cell.2024.05.035."
COMMENT     Sequence Label: pET28_Human_SAT1_Nterm_His
FEATURES             Location/Qualifiers
     source          1..5805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..40
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(34..225)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1034..2116)
                     /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     2078..2097
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2117..2194)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2400..2419
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2503..2522
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2503..2521
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2522..2546
                     /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             2561..2583
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2602..2619
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2629..2646
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3172..3189
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3242..3260)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3256..3303
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3340..3795
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3427..3446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3637..3658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3887..4702)
                     /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(3887..4702)
                     /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     4610..4629
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4694..4713
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4824..5412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5313..5332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5566..5583
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5598..5737
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5663..5682)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
       61 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
      121 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
      181 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
      241 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
      301 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
      361 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
      421 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
      481 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
      541 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
      601 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
      661 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
      721 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
      781 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
      841 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
      901 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
      961 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     1021 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1081 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     1141 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     1201 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     1261 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     1321 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     1381 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     1441 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     1501 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     1561 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     1621 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     1681 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     1741 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     1801 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     1861 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     1921 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     1981 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     2041 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     2101 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     2161 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     2221 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     2281 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     2341 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     2401 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     2461 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag
     2521 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
     2581 agatatacca tgggcagcag ccatcatcat catcatcaca gcagcggcct ggtgccgcgc
     2641 ggcagccata tggccaaatt tgttattcgt ccggcaaccg cagcagattg tagcgatatt
     2701 ctgcgtctga ttaaagaact ggccaaatac gagtatatgg aagaacaggt tatcctgacc
     2761 gaaaaagatc tgctggaaga tggttttggt gaacatccgt tttatcattg tctggttgca
     2821 gaagttccga aagaacattg gacaccggaa ggtcatagca ttgttggttt tgccatgtac
     2881 tatttcacct atgatccgtg gattggcaaa ctgctgtatc tggaagattt ctttgtgatg
     2941 agcgattatc gcggttttgg tattggttcc gaaattctga aaaatctgag ccaggttgca
     3001 atgcgttgtc gttgtagcag catgcatttt ctggtggccg aatggaatga accgagcatt
     3061 aatttctata aacgtcgtgg tgcaagcgat ctgagcagcg aagaaggttg gcgtctgttt
     3121 aaaatcgata aagaatacct gctgaaaatg gccaccgaag aataactcga gcaccaccac
     3181 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     3241 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     3301 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     3361 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3421 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3481 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3541 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3601 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3661 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3721 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3781 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3841 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     3901 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     3961 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     4021 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     4081 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     4141 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     4201 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     4261 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     4321 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     4381 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     4441 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     4501 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     4561 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     4621 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     4681 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     4741 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4801 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4861 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4921 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     4981 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5041 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5101 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5161 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5221 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5281 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5341 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5401 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5461 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5521 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     5581 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     5641 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     5701 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     5761 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacag
//
