LOCUS       sequence-434031-        3269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heterologous expression of PA1377 under endogenous speG E. coli 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3269)
  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 3269)
  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_speGpromoter_PA1377
FEATURES             Location/Qualifiers
     source          1..3269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      433..1021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     922..941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1175..1192
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      2355..2441
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2533..2560
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2579..2670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2671..2739,2740..3269,1..262)
                     /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(2889..2908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
       61 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
      121 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
      181 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
      241 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
      301 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
      361 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
      421 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
      481 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      541 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
      601 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      661 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      721 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      781 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      841 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      901 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      961 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1021 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     1081 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     1141 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     1201 agagcgcctg atgcggtatt ttctccaaaa tagtaacgtt gttttaatca tctttgtctc
     1261 ttattgacat catgtatagt tatagggcga cataatatca tcaatataaa caccctcctg
     1321 gtacgttttg cgtccgcagt ggatggtgta gaatcacctt taatcattca tacagggaat
     1381 gaattatgaa aatcactctc agcaaacgaa tcggcctgct cgctattctg ctgccttgcg
     1441 cactggcatt gagcacaact gttcatgccg aaactaacaa actggtgatt gagtctggcg
     1501 acagtgcaca aagccgccag cacgccgcta tggaaaaaga gcaatggaat gacacgcgca
     1561 atctgcgcca gaaagtgaat aaacgcactg aaaaagagtg ggataaagcc gacgccgctt
     1621 ttgataaccg cgataaatgt gagcaaagcg ccaacatcaa tgcctactgg gagcccaata
     1681 ctttgcgctg cctggaccgt cgaactggcc gcgttattac cccctaacct gttattgatt
     1741 taaggaatgt aaggacacgt tatgaccgca gaaagcccga ccattcgtct ggaacgttat
     1801 agcgaacgtc atgttgaagg tctgaccgca ctgtataatg atccggcagt tgcacgtcag
     1861 gttctgcaga tgccgtatca gagcgttgaa cagcgtcgta aacgtctgca tgatagcgca
     1921 gatgatgatc gtctgctgat tctggttgca ctgcatcagg gtgatgttat tggtagcgca
     1981 agcctggaac agcatccgcg tattcgtcgt agccatagcg gtagcattgg tatgggtgtt
     2041 gcagttgcat ggcaaggtaa aggtgttggt agccgtctgc tgggtgaact gctggatatt
     2101 gcagataatt ggatgaatct gcgtcgtgtt gaactgaccg tttataccga taatgcaccg
     2161 gcactggcac tgtatcgtaa atttggtttt gaaaccgaag gtgagatgcg tgattatgca
     2221 gttcgtgatg gtcgttttgt tgatgtttat agcatggcac gtctgcgtcg cgtggaaggt
     2281 cgtgttggtg aataactgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     2341 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     2401 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     2461 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     2521 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     2581 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2641 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2701 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2761 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2821 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2881 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     2941 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3001 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3061 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3121 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3181 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3241 ctattaactg gcgaactact tactctagc
//
