LOCUS       sequence-217605-       11760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of mariner-based transposon mutant libraries in 
            Enterococcus faecium.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11760)
  AUTHORS   Zhang X, de Maat V, Guzman Prieto AM, Prajsnar TK, Bayjanov JR, de 
            Been M, Rogers MRC, Bonten MJM, Mesnage S, Willems RJL, van Schaik W
  TITLE     RNA-seq and Tn-seq reveal fitness determinants of 
            vancomycin-resistant Enterococcus faecium during growth in human 
            serum.
  JOURNAL   BMC Genomics. 2017 Nov 21;18(1):893. doi: 10.1186/s12864-017-4299-9.
  PUBMED    29162049
REFERENCE   2  (bases 1 to 11760)
  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: "BMC 
            Genomics."; date: "2017-11-21"; volume: "18(1):893. doi"; pages: " 
            10.1186/s12864-017-4299-9"
COMMENT     Sequence Label: pGPA1
FEATURES             Location/Qualifiers
     source          1..11760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1051..1062)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(4372..4390)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4391..4407
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4391..4407
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6281..6299
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8241..8259)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8346..8363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8517..9105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8597..8616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattctgta caattgctag cgtacggccg tcgacgtccg gatcctcgag ctctagatct
       61 taagacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      121 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      181 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      241 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
      301 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
      361 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
      421 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
      481 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
      541 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
      601 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccggggga tcgttaaatt
      661 tatactgcaa tcggatgcga ttattgaata aaagatatga gagatttatc taatttcttt
      721 tttcttgtaa aaaaagaaag ttcttaaagg ttttatagtt ttggtcgtag agcacacggt
      781 ttaacgactt aattacgaag taaataagtc tagtgtgtta gactttatga aatctatata
      841 cgtttatata tatttattat ccgatttttt attaaaacgt ctcaaaatcg tttctgagac
      901 gttttagcgt ttatttcgtt tagttatcgg cataatcgtt aaaacaggcg ttatcgtagc
      961 gtaaaagccc ttgagcgtag cgtggctttg cagcgaagat gttgtctgtt agattatgaa
     1021 agccgatgac tgaatgaaat aataagcgca gcgcccttct atttcggttg gaggaggctc
     1081 aagggagtat gagggaatga aattccctca tgggtttgat tttaaaaatt gcttgcaatt
     1141 ttgccgagcg gtagcgctgg aaaatttttg aaaaaaattt ggaatttgga aaaaaatggg
     1201 gggaaaggaa gcgaattttg cttccgtact acgacccccc attaagtgcc gagtgccaat
     1261 ttttgtgcca aaaacgctct atcccaactg gctcaagggt ttaaggggtt tttcaatcgc
     1321 caacgaatcg ccaacgtttt cgccaacgtt ttttataaat ctatatttaa gtagctttat
     1381 tgttgttttt atgattacaa agtgatacac taactttata aaattatttg attggagttt
     1441 tttaaatggt gatttcagaa tcgaaaaaaa gagttatgat ttctctgaca aaagagcaag
     1501 ataaaaaatt aacagatatg gcgaaacaaa aaggtttttc aaaatctgcg gttgcggcgt
     1561 tagctataga agaatatgca agaaaggaat cagaataaaa aaaataagcg aaagctcgcg
     1621 tttttagaag gatacgagtt ttcgctactt gtttttgata aggtaattat atcatggcta
     1681 ttaaaaatac taaagctaga aattttggat ttttattata tcctgactca attcctaatg
     1741 attggaaaga aaaattagag agtttgggcg tatctatggc tgtcagtcct ttacacgata
     1801 tggacgaaaa aaaagataaa gatacatgga ataatagtaa tattatacaa aatggaaagc
     1861 actataaaaa accacactat cacgttatat atattgcacg aaatcctgta acaatagaaa
     1921 gcgttaggaa caagattaag cgaaaattgg ggaatagttc agttgctcat gttgagatac
     1981 ttgattatat caaaggttca tatgaatatt tgactcatga atcaaaggac gctattgcta
     2041 agaataaaca tatatacgac aaaaaagata ttttgaacat taatgatttt gatattgacc
     2101 gctatataac acttgatgaa agccaaaaaa gagaattgaa gaatttactt ttagatatag
     2161 tggatgacta taatttggta aatacaaaag atttaatggc ttttattcgc cttaggggag
     2221 cggagtttgg aattttaaat acgaatgatg taaaagatat tgtttcaaca aactctagcg
     2281 cctttagatt atggtttgag ggcaattatc agtgtggata tagagcaagt tatgcaaagg
     2341 ttcttgatgc tgaaacgggg gaaataaaat gacaaacaaa gaaaaagagt tatttgctga
     2401 aaatgaggaa ttaaaaaaag aaattaagga cttaaaagag cgtattgaaa gatacagaga
     2461 aatggaagtt gaattaagta caacaataga tttattgaga ggagggatta ttgaataaat
     2521 aaaagccccc tgacgaaagt cgaagggggt ttttattttg gtttgatgtt gcgattaata
     2581 gcaatacaat tgcaataaac aaaatgatcc tctagagtcg atacaaattc ctcgtaggcg
     2641 ctcgggaccc ctatctagcg aacttttaga aaagatataa aacatcagag tatggacagt
     2701 tgcggatgta cttcagaaaa gattagatgt ctaaaaagct tgtagttaaa gctttttaga
     2761 catctaaatc taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc
     2821 aatcaggctt gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgataa
     2881 ttcgatgatc tggagctgta atataaaaac cttcttcaac taacggggca ggttagtgac
     2941 attagaaaac cgactgtaaa aagtacagtc ggcattatct catattataa aagccagtca
     3001 ttaggcctat ctgacaattc ctgaatagag ttcataaaca atcctgcatg ataaccatca
     3061 caaacagaat gatgtacctg taaagatagc ggtaaatata ttgaattacc tttattaatg
     3121 aattttcctg ctgtaataat gggtagaagg taattactat tattattgat atttaagtta
     3181 aacccagtaa atgaagtcca tggaataata gaaagagaaa aagcattttc aggtataggt
     3241 gttttgggaa acaatttccc cgaaccatta tatttctcta catcagaaag gtataaatca
     3301 taaaactctt tgaagtcatt ctttacagga gtccaaatac cagagaatgt tttagataca
     3361 ccatcaaaaa ttgtataaag tggctctaac ttatcccaat aacctaactc tccgtcgcta
     3421 ttgtaaccag ttctaaaagc tgtatttgag tttatcaccc ttgtcactaa gaaaataaat
     3481 gcagggtaaa atttatatcc ttcttgtttt atgtttcggt ataaaacact aatatcaatt
     3541 tctgtggtta tactaaaagt cgtttgttgg ttcaaataat gattaaatat ctcttttctc
     3601 ttccaattgt ctaaatcaat tttattaaag ttcatttgat atgcctccta aatttttatc
     3661 taaagtgaat ttaggaggct tacttgtctg ctttcttcat tagaatcaat ccttttttaa
     3721 aagtcaatat tactgtaaca taaatatata ttttaaaaat atcccacttt atccaatttt
     3781 cgtttgttga actaatgggt gctttagttg aagaataaaa gaccacatta aaaaatgtgg
     3841 tcttttgtgt ttttttaaag gatttgagcg tagcgaaaaa tccttttctt tcttatcttg
     3901 ataataaggg taactattgc cgatcaagct atcttctata taaaagatat attatcttat
     3961 cagtattgtc aatatattca aggcaatctg cctcctcatc ctcttcatcc tcttcgtctt
     4021 ggtagctttt taaatatggc gcttcataga gtaattctgt aaaggtccaa ttctcgtttt
     4081 catacctcgg tataatctta cctatcacct caaatggttc gctgggttta tcgacctgca
     4141 ggtcgactct agaggatccc ccggtgcggg cctcttcgct attacgccag cttaagaaag
     4201 tcgaaggggg tttttatttt ggtttgatgt tgcgattaat agcaatacaa ttgcaataaa
     4261 caaaatgatc ctctagagca tccgcggtaa caggttggat gataagtccc cggtctaaca
     4321 aagaaaaaca catttttttg ttaaaattcg ttttagatct atacccgccc tcctatagtg
     4381 agtcgtatta caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc
     4441 gatccccctt ttcatatatt atatagtcaa tctttataag tccttttata aatttctttt
     4501 ctaccatttt cgataaattc ctgtttaata tttttaattc cataaacaat agtttcaata
     4561 ggataatatt cttcaactat atcttgatat tcttttgctt tctcaatatc tatatttcca
     4621 tacattctta atatatcttc tccaaaattt gttcctattt cttcttcact atcttcaagt
     4681 aagtatataa aatcacaata ttcatctata attccagaat ctccaaaatc aattatccca
     4741 gttaatctat tattgccatc taacaataga tgattacaac taaaatcatt atggcataaa
     4801 cactttttac cctcaaaaac tgttgttgca tttagtcttt ccataaaact ttctatataa
     4861 tctttttcta tatcagttaa atcattataa atagtttcac gcaacaatat atactcttct
     4921 aatacatttt gtttattatc aatagtacat tcactaatat ctgtataatc taaaccgtgc
     4981 atttgtctta aaaaactggc aatatctcgt tttaacaaat tttgttcttc ttctgacata
     5041 gtagaataaa tttctggtgt taaaaaagtt cctttaattt ctttataacc tagtatagat
     5101 aattcatcac taatatacga atattcaata ttaggaattt ttacattagt ttctaaattt
     5161 gtatttaaaa aattatatat tgctttttct tttgcataac cttttttctt attagtacta
     5221 aattttgttt taaaaatgta ttcattatta actaaatatg ccacactatc ataaccacta
     5281 ccgattattt caatactatc tactttgaaa ttatcaaagt aatgctcaat taaatatttc
     5341 attgccttaa catttgtggc attatcatca tatctatatt ccattaaata acaatcttct
     5401 tttttgccct cgtgtaattc atgttctggc aaatcttcaa taattctaaa accagatctt
     5461 tggtatgccc ttattgctct tggattattt ttatgagggt ctaaaataac tgcattagca
     5521 tttctttctt ttttcaaaaa ttcagaaatc aatttaatat atcttgtacc aattccttta
     5581 ctccaataat tcggctctcc tataaattga tccataccat agactatctc atcagttttt
     5641 ggataatgat aatcagtata taactcatca tacattttat atatttgtcc atatccaata
     5701 ggaacattgt tatattcaat aattactcta aaaacttcat cttcccaagg ctctgtataa
     5761 tgttttttta atgattctaa tgtatatttt ttatctctac caccataaaa ttctaatact
     5821 ctttcatcag ttaaccattt taacatcaaa ggaaaatcat catctattaa agttcttata
     5881 catatttcat tttcaactat attcatttat ttatcacctt tttcataatc atatacatat
     5941 actatttcat ctttataatc atttttacca cctaattttt catatacatg gcaaggggaa
     6001 ttgactctag aggatccccg ggtaccgagc tcgccccgcg ctgcctatgc cttgccccct
     6061 gaaatcctta catacggcga tatcttctat ataaaagata tattatctta tcagtattgt
     6121 caatatattc aaggcaatct gcctcctcat cctcttcatc ctcttcgtct tggtagcttt
     6181 ttaaatatgg cgcttcatag agtaattctg taaaggtcca attctcgttt tcatacctcg
     6241 gtataatctt acctatcacc tcaaatggtt cgctgggttt taatacgact cactatagga
     6301 gggcgggaat catttgaagg ttggtactat ataaaaataa tatgcattta atactagcga
     6361 cgccatctat gtgtcagacc ggggacttat catccaacct gttaccgcgg agataggtta
     6421 tgcaagattt ttatatggaa atagcatgaa acaaaaatga attaactata attactaaat
     6481 tcttatttac agtcaatcag atttttgatt aaaaggagta gagaagtcga attcccgtga
     6541 atcgaaaacc gctacggatc agatctagtc ttataactat actgacaata gaaacattaa
     6601 caaatctaaa acagtcttaa ttctatcttg agaaagtatt ggtaataata ttattgtcga
     6661 taacgcgagc ataataaacg gctctgatta aattctgaag tttgttagat acaatgattt
     6721 cgttcgaagg aactacaaaa taaattataa ggaggcactc accatggaaa aaaaggaatt
     6781 tcgtgttttg ataaaatact gttttctgaa gggaaaaaat acagtggaag caaaaacttg
     6841 gcttgataat gagtttccgg actctgcccc agggaaatca acaataattg attggtatgc
     6901 aaaattcaag cgtggtgaaa tgagcacgga ggacggtgaa cgcagtggac gcccgaaaga
     6961 ggtggttacc gacgaaaaca tcaaaaaaat ccacaaaatg attttgaatg accgtaaaat
     7021 gaagttgatc gagatagcag aggccttaaa gatatcaaag gaacgtgttg gtcatatcat
     7081 tcatcaatat ttggatatgc ggaagctctg tgcgaaatgg gtgccgcgcg agctcacatt
     7141 tgaccaaaaa caacgacgtg ttgatgattc taagcggtgt ttgcagctgt taactcgtaa
     7201 tacacccgag tttttccgtc gatatgtgac aatggatgaa acatggctcc atcactacac
     7261 tcctgagtcc aatcgacagt cggctgagtg gacagcgacc ggtgaaccgt ctccgaagcg
     7321 tggaaagact caaaagtccg ctggcaaagt aatggcctct gttttttggg atgcgcatgg
     7381 aataattttt atcgattatc ttgagaaggg aaaaaccatc aacagtgact attatatggc
     7441 gttattggag cgtttgaagg tcgaaatcgc ggcaaaacgg ccccacatga agaagaaaaa
     7501 agtgttgttc caccaagaca acgcaccgtg ccacaagtca ttgagaacga tggcaaaaat
     7561 tcatgaattg ggcttcgaat tgcttcccca cccgccgtat tctccagatc tggcccccag
     7621 cgactttttc ttgttctcag acctcaaaag gatgctcgca gggaaaaaat ttggctgcaa
     7681 tgaagaggtg atcgccgaaa ctgaggccta ttttgaggca aaaccgaagg agtactacca
     7741 aaatggtatc aaaaaattgg aaggtcgtta taatcgttgt atcgctcttg aagggaacta
     7801 tgttgaataa taaaaacgaa ttttcacaaa aaaatgtgtt tttctttgtt agaccggatc
     7861 tgcaggcatg cggtaccact agttctagag agctcaagct ttctttgaac caaaattaga
     7921 aaaccaaggc ttgaaacgtt caattgaaat ggcaattaaa caaattacag cacgtgttgc
     7981 tttgattgat agccaaaaag cagcagttga taaagcaatt actgatattg ctgaaaaatt
     8041 gtaatttata aataaaaatc accttttaga ggtggttttt ttatttataa attattcgtt
     8101 tgatttcgct ttcgatagaa caatcaaagc gagaataagg aagataaatc ccataagggc
     8161 gggagcagaa tgtccgagac taatgtaaat ttgtccacca attaaaggac cgataacgcg
     8221 agcttctcga ggccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac
     8281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     8341 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     8401 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     8461 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     8521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     8581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8641 cctgttccga ccctgccgct taccggatac ctgcccgcct ttctcccttc gggaagcgtg
     8701 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     8881 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     8941 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     9001 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     9061 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     9121 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     9181 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     9241 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     9301 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     9361 ataactacga tacgggaggg cttaccatct ggccccggtg ctgcaatgat accgcgagac
     9421 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     9481 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     9541 agagtaagta gttcgccagt taatagtttg ccgaaacaaa aaaagtaatc cttagagatt
     9601 actaaattac ttttttattt ttaggataac ttctgcccca cctttttgag gattactgag
     9661 aattaagttt ccttgatgtt ttaaagctac accttgagca aaagatagtc caatcccata
     9721 gtgtttccca ctacgtccag tatcttctgt gaaaaatagt tttccagcat ttttttttgc
     9781 ttgttcagaa aaaggatgac cattattcca gatttcaaaa tagagatatt tttcatcatc
     9841 ataaatactc aaaccgacta ttttttcacc ctctttagca tactgacagg cattaacaaa
     9901 gatattgata agtgctcgac ttaaagctaa tgtatttccg taaaaagtgg ttaaatctgt
     9961 ttttttaact agctgataat ccacttgata agttgttgac aactcttcca actcaactga
    10021 taaatctatc aaaaaatcct ccagggagat tatcgctttg taatcatttt catcattcaa
    10081 aagttttgta taactaatca ttgtgttaaa atacttgtca aaaacagtca aactattttt
    10141 cattgactca ataaaatcag cttgttgttc atttacttca gtcatctcta gcagttcaat
    10201 atttcctttt aatactgtta aaggtgtctt aacatcatgt gaaagtgcag caacctggaa
    10261 ggataaatca tgtttttcat gacgctccgc ttcaataagg aaagctaact ccttactttt
    10321 cgaatataag ttattcagaa cctgatcaaa ttcaataatt tttgattcct cttgttcagg
    10381 aaaagtagtt atttccccca tcttcaatgc aatcttttga acggcttgaa aattcttaga
    10441 aaattctcta atgagatgat agacagaaaa aataatgagt attatttcac caagaaaaaa
    10501 gaaaaggtaa gtaaattgat tataagaaat tgaacgaagc gtatgatttg taaattcagg
    10561 catactgttt tgtcttaaaa caactgaaaa atgcttattg cttgaataag tataagtaat
    10621 ttctccttct gtaatattat gtgaagactg ttttgaagct acaaaactag gtacatccga
    10681 cttgacataa tgtcctccca aaattttact tgtatcattg tcaaaaataa tataatcata
    10741 tgggtagtgt ttcatatcac gttcgatatc agaaatagtt aaattatctg aatctaaaga
    10801 taaacgaata gtttctgaac cactgatttg tccaatttgt cgtagaaaga aagtcaagcc
    10861 cagtaacaac aggataagta gacaagtacc tatgataatt tcaatgacag cttgccatat
    10921 ccgtcgacgc attgaatatt ttttacccat gccactgata cccaagtccc cgaaccgttt
    10981 ttatcggatt aatatcgtat ggtgcaaact tactcctaat ctgataaata tattccgaga
    11041 ttgaccgaaa aagtgcatta gaatattcat catatacgtc atcataaata tcctctctcg
    11101 tataaacttt agaagttcgt tgtgatagta attcaagaat atcgtattcg cgacaagtca
    11161 gtggaattgt ttgaccattg acacagactc gcctttcttc taaataaaat gtaattggtc
    11221 ctaaatctct acgaatctct gaaaaaacat gaactgcatg tttattgcgt tcctctcgct
    11281 ttatatttgc ttccactttt gcaaccaact gtttaaggct aaaaggctta gtaatatagt
    11341 catccccacc aatacctaag ccgtttataa tatcctcttc tgtatctttc gcactaacaa
    11401 agataattgg agttgatatt tctctgcgaa tccttttaca aatttctgtc ccttcaatat
    11461 ttgacatcat gatatctaac aaaatcaaat caaatccctg aaaatcagta atatccaagg
    11521 gaagtgaaat gttttgatgc gtcgcaactt catagtttct catttctaat gctgtcttca
    11581 ttaattttaa aatttcctga tcatcatcaa ctattaaaat tttatacacc actttacctc
    11641 cgattgttta gtactcatta cttcattata ccattcattc tgaagaaata taatctatca
    11701 atttttagtc tttcttttca atataccaag gattgagata cacactatac ctaaagccgg
//
