LOCUS       sequence-437211-        3296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heterologous expression under endogenous speG E. coli promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3296)
  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 3296)
  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: pTrcHis_VectorA_short_speG_genomic
FEATURES             Location/Qualifiers
     source          1..3296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      303..891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     792..811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1045..1062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      2252..2338
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2430..2457
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2476..2567
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2568..2636,2637..3296,1..132)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2786..2805)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
       61 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
      121 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
      181 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      241 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      301 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      361 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      421 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
      481 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      541 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      601 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      661 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      721 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      781 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      841 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      901 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      961 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     1021 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     1081 atgcggtatt ttctccaaaa tagtaacgtt gttttaatca tctttgtctc ttattgacat
     1141 catgtatagt tatagggcga cataatatca tcaatataaa caccctcctg gtacgttttg
     1201 cgtccgcagt ggatggtgta gaatcacctt taatcattca tacagggaat gaattatgaa
     1261 aatcactctc agcaaacgaa tcggcctgct cgctattctg ctgccttgcg cactggcatt
     1321 gagcacaact gttcatgccg aaactaacaa actggtgatt gagtctggcg acagtgcaca
     1381 aagccgccag cacgccgcta tggaaaaaga gcaatggaat gacacgcgca atctgcgcca
     1441 gaaagtgaat aaacgcactg aaaaagagtg ggataaagcc gacgccgctt ttgataaccg
     1501 cgataaatgt gagcaaagcg ccaacatcaa tgcctactgg gagcccaata ctttgcgctg
     1561 cctggaccgt cgaactggcc gcgttattac cccctaacct gttattgatt taaggaatgt
     1621 aaggacacgt tatgccaagc gcccacagtg ttaagctacg cccgctggag cgtgaagatt
     1681 tacgctatgt acatcaactc gacaataacg ccagtgtgat gcgttactgg tttgaggaac
     1741 cctacgaagc ctttgttgaa ctctctgatc tgtatgataa gcatattcac gatcagagcg
     1801 aacggcgctt tgtggtggaa tgtgacggcg aaaaagccgg tctggtggag ctggtggaaa
     1861 ttaaccatgt tcatcgccgc gcagaatttc agataattat ctccccggag tatcagggga
     1921 aaggtctggc aacccgtgcc gccaaattag caatggacta tggctttacc gttctcaatc
     1981 tctataagct gtatctgatc gttgataaag agaatgaaaa agcgattcac atttaccgca
     2041 agcttggctt ttcggttgaa ggtgaattga tgcacgagtt ctttattaat ggtcaatatc
     2101 gtaatgccat tcgcatgtgt atattccagc atcagtatct ggcagagcac aaaacaccgg
     2161 gtcagactct cctgaagccg accgcacaat agccgtagcg ccgatggtag tgtggggtct
     2221 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2281 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     2341 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     2401 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     2461 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2521 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2581 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2641 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2701 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2761 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     2821 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2881 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2941 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3001 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3061 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3121 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3181 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3241 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatc
//
