LOCUS       sequence-434041-        5742 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 5742)
  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 5742)
  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_Cterm_His
FEATURES             Location/Qualifiers
     source          1..5742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..63
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(57..248)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1057..2139)
                     /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     2101..2120
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2140..2217)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2423..2442
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2526..2545
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2526..2544
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2545..2569
                     /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             2584..2606
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3132..3149
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3202..3220)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3216..3263
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3300..3755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3387..3406)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3597..3618
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3847..4662)
                     /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(3847..4662)
                     /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     4570..4589
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4654..4673
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4784..5372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5273..5292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5526..5543
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5558..5697
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5623..5642)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
       61 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
      121 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
      181 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
      241 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
      301 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
      361 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
      421 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
      481 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
      541 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
      601 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
      661 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
      721 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
      781 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
      841 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
      901 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
      961 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     1021 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     1081 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1141 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     1201 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     1261 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     1321 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     1381 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     1441 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     1501 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     1561 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     1621 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     1681 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     1741 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     1801 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     1861 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     1921 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     1981 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     2041 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     2101 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     2161 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     2221 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     2281 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     2341 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     2401 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     2461 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     2521 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
     2581 aattttgttt aactttaaga aggagatata ccatggccaa atttgttatt cgtccggcaa
     2641 ccgcagcaga ttgtagcgat attctgcgtc tgattaaaga actggccaaa tacgagtata
     2701 tggaagaaca ggttatcctg accgaaaaag atctgctgga agatggtttt ggtgaacatc
     2761 cgttttatca ttgtctggtt gcagaagttc cgaaagaaca ttggacaccg gaaggtcata
     2821 gcattgttgg ttttgccatg tactatttca cctatgatcc gtggattggc aaactgctgt
     2881 atctggaaga tttctttgtg atgagcgatt atcgcggttt tggtattggt tccgaaattc
     2941 tgaaaaatct gagccaggtt gcaatgcgtt gtcgttgtag cagcatgcat tttctggtgg
     3001 ccgaatggaa tgaaccgagc attaatttct ataaacgtcg tggtgcaagc gatctgagca
     3061 gcgaagaagg ttggcgtctg tttaaaatcg ataaagaata cctgctgaaa atggccaccg
     3121 aagaactcga gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg
     3181 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     3241 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga
     3301 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3361 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3421 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3481 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3541 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3601 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3661 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3721 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     3781 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     3841 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     3901 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     3961 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     4021 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     4081 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     4141 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     4201 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     4261 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     4321 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     4381 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     4441 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     4501 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     4561 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     4621 tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     4681 gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt
     4741 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4801 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4861 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4921 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4981 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5041 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5101 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5161 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5221 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5281 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5341 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5401 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5461 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5521 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     5581 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     5641 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     5701 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gt
//
