LOCUS       sequence-437213-        3290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heterologous expression of PA1472 under endogenous speG E. coli 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3290)
  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 3290)
  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_PA1472
FEATURES             Location/Qualifiers
     source          1..3290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1004..1090
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1182..1209
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1228..1319
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1320..1388,1389..2180)
                     /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(1538..1557)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2351..2939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2840..2859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3093..3110
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagggaatga attatgaaaa tcactctcag caaacgaatc ggcctgctcg ctattctgct
       61 gccttgcgca ctggcattga gcacaactgt tcatgccgaa actaacaaac tggtgattga
      121 gtctggcgac agtgcacaaa gccgccagca cgccgctatg gaaaaagagc aatggaatga
      181 cacgcgcaat ctgcgccaga aagtgaataa acgcactgaa aaagagtggg ataaagccga
      241 cgccgctttt gataaccgcg ataaatgtga gcaaagcgcc aacatcaatg cctactggga
      301 gcccaatact ttgcgctgcc tggaccgtcg aactggccgc gttattaccc cctaacctgt
      361 tattgattta aggaatgtaa ggacacgtta tgaactttgt ttttccgcat ccgctggata
      421 caccgcgtct gcgtctgcgc acctttcgtg aagaagatgc agcaccgctg tttgcaatga
      481 tgagcgatcc ggaagttatg cgttattgga atacccctcc gtggaccaca ccggcacagg
      541 cacgtgaagc aattctgcgt gatagccagg cactggttga tggtgaatat ctgaatctgg
      601 caattgaacg tcgtgaagat ggtcagctgc tgggtagctg tattctgttt cattttgaaa
      661 aaggtagccg tcgtgcagaa ctgggttatt gtctggcacg tgcagcacaa ggtcgtggtt
      721 atatgggtga agcactgcgt cgtctgctgg catttgcatt tgatgaaatt gatctgaatc
      781 gtctggaagc agaaatcgat ccgcgtaatc gtccgagcgc agcaagcctg gaacgtctgg
      841 gttttcgtca agaaggcctg ctggcacagc gttggattgt tagcggtgaa gttagcgata
      901 gcgcactgta tggactgctg gcagaacatt ggcgtaatcg ctaaccgtag cgccgatggt
      961 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     1021 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     1081 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     1141 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     1201 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
     1261 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1321 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1381 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1441 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1501 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1561 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1621 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1681 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1741 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1801 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     1861 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     1921 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     1981 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2041 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2101 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2161 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2221 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2281 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2341 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2401 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2461 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2521 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2581 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2641 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2701 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2761 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2821 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2881 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2941 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3001 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3061 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3121 agcgcctgat gcggtatttt ctccaaaata gtaacgttgt tttaatcatc tttgtctctt
     3181 attgacatca tgtatagtta tagggcgaca taatatcatc aatataaaca ccctcctggt
     3241 acgttttgcg tccgcagtgg atggtgtaga atcaccttta atcattcata
//
