LOCUS       sequence-434029-        3245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heterologous expression of PA4114 under endogenous speG E. coli 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3245)
  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 3245)
  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_PA4114
FEATURES             Location/Qualifiers
     source          1..3245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      300..888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     789..808
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1042..1059
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      2198..2284
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2376..2403
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2422..2513
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2514..2582,2583..3245,1..129)
                     /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(2732..2751)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
       61 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
      121 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
      181 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
      241 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
      301 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
      361 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
      421 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
      481 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      541 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      601 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      661 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      721 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      781 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      841 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      901 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      961 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     1021 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     1081 cggtattttc tccaaaatag taacgttgtt ttaatcatct ttgtctctta ttgacatcat
     1141 gtatagttat agggcgacat aatatcatca atataaacac cctcctggta cgttttgcgt
     1201 ccgcagtgga tggtgtagaa tcacctttaa tcattcatac agggaatgaa ttatgaaaat
     1261 cactctcagc aaacgaatcg gcctgctcgc tattctgctg ccttgcgcac tggcattgag
     1321 cacaactgtt catgccgaaa ctaacaaact ggtgattgag tctggcgaca gtgcacaaag
     1381 ccgccagcac gccgctatgg aaaaagagca atggaatgac acgcgcaatc tgcgccagaa
     1441 agtgaataaa cgcactgaaa aagagtggga taaagccgac gccgcttttg ataaccgcga
     1501 taaatgtgag caaagcgcca acatcaatgc ctactgggag cccaatactt tgcgctgcct
     1561 ggaccgtcga actggccgcg ttattacccc ctaacctgtt attgatttaa ggaatgtaag
     1621 gacacgttat gattgaactg cgtaccatta cacgtgatga ttgggaaacc tgtattgatc
     1681 tgaaagttgc acgtcatcag gcacattttg ttgcaagcaa tctgtatagc ctggcacaga
     1741 gccgttttct gcctggtttt cgtgcagttg gtattcatca tggtggtcgt atggttggtt
     1801 ttgcactgta tggtccggat gcagatgatg gtcattattg gctgtatcgt ctgatgattg
     1861 atcgtcgcta tcaaggtaaa ggtcatggtc gtgcagcact gcagcgtatt attgcaagcg
     1921 ttcgtcgtac ccgtcgtcgt agcgatcgcc tgatggttgc atatgccgtt gataatgatg
     1981 ttgcccgtca gctgtaccgt gatgcaggtt ttgttgaaca aggtctggca ccgtggggtg
     2041 aaaccatggc atgtctgatg ctgccggaac cgagccatcg tagcggtaaa gttgccggtg
     2101 caccaggtgg tcgcgcagcc catggtgttc cggcataacc gtagcgccga tggtagtgtg
     2161 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     2221 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     2281 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     2341 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
     2401 ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc
     2461 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2521 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2581 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2641 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2701 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     2761 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2821 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2881 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2941 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3001 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3061 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3121 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3181 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3241 tcatt
//
