LOCUS       Exported               11760 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Generation of mariner-based transposon mutant libraries in 
            Enterococcus faecium.
ACCESSION   .
VERSION     .
KEYWORDS    pGPA1
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11760
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      2996..3584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3485..3504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3738..3755
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3842..3861
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3842..3860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5802..5820)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7694..7710)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7694..7710)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7711..7729
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             11039..11050
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
       61 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
      121 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
      181 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
      241 aaatccctta acgtgagttt tcgttccact gagcgtctta agatctagag ctcgaggatc
      301 cggacgtcga cggccgtacg ctagcaattg tacagaattc ccggctttag gtatagtgtg
      361 tatctcaatc cttggtatat tgaaaagaaa gactaaaaat tgatagatta tatttcttca
      421 gaatgaatgg tataatgaag taatgagtac taaacaatcg gaggtaaagt ggtgtataaa
      481 attttaatag ttgatgatga tcaggaaatt ttaaaattaa tgaagacagc attagaaatg
      541 agaaactatg aagttgcgac gcatcaaaac atttcacttc ccttggatat tactgatttt
      601 cagggatttg atttgatttt gttagatatc atgatgtcaa atattgaagg gacagaaatt
      661 tgtaaaagga ttcgcagaga aatatcaact ccaattatct ttgttagtgc gaaagataca
      721 gaagaggata ttataaacgg cttaggtatt ggtggggatg actatattac taagcctttt
      781 agccttaaac agttggttgc aaaagtggaa gcaaatataa agcgagagga acgcaataaa
      841 catgcagttc atgttttttc agagattcgt agagatttag gaccaattac attttattta
      901 gaagaaaggc gagtctgtgt caatggtcaa acaattccac tgacttgtcg cgaatacgat
      961 attcttgaat tactatcaca acgaacttct aaagtttata cgagagagga tatttatgat
     1021 gacgtatatg atgaatattc taatgcactt tttcggtcaa tctcggaata tatttatcag
     1081 attaggagta agtttgcacc atacgatatt aatccgataa aaacggttcg gggacttggg
     1141 tatcagtggc atgggtaaaa aatattcaat gcgtcgacgg atatggcaag ctgtcattga
     1201 aattatcata ggtacttgtc tacttatcct gttgttactg ggcttgactt tctttctacg
     1261 acaaattgga caaatcagtg gttcagaaac tattcgttta tctttagatt cagataattt
     1321 aactatttct gatatcgaac gtgatatgaa acactaccca tatgattata ttatttttga
     1381 caatgataca agtaaaattt tgggaggaca ttatgtcaag tcggatgtac ctagttttgt
     1441 agcttcaaaa cagtcttcac ataatattac agaaggagaa attacttata cttattcaag
     1501 caataagcat ttttcagttg ttttaagaca aaacagtatg cctgaattta caaatcatac
     1561 gcttcgttca atttcttata atcaatttac ttaccttttc ttttttcttg gtgaaataat
     1621 actcattatt ttttctgtct atcatctcat tagagaattt tctaagaatt ttcaagccgt
     1681 tcaaaagatt gcattgaaga tgggggaaat aactactttt cctgaacaag aggaatcaaa
     1741 aattattgaa tttgatcagg ttctgaataa cttatattcg aaaagtaagg agttagcttt
     1801 ccttattgaa gcggagcgtc atgaaaaaca tgatttatcc ttccaggttg ctgcactttc
     1861 acatgatgtt aagacacctt taacagtatt aaaaggaaat attgaactgc tagagatgac
     1921 tgaagtaaat gaacaacaag ctgattttat tgagtcaatg aaaaatagtt tgactgtttt
     1981 tgacaagtat tttaacacaa tgattagtta tacaaaactt ttgaatgatg aaaatgatta
     2041 caaagcgata atctccctgg aggatttttt gatagattta tcagttgagt tggaagagtt
     2101 gtcaacaact tatcaagtgg attatcagct agttaaaaaa acagatttaa ccacttttta
     2161 cggaaataca ttagctttaa gtcgagcact tatcaatatc tttgttaatg cctgtcagta
     2221 tgctaaagag ggtgaaaaaa tagtcggttt gagtatttat gatgatgaaa aatatctcta
     2281 ttttgaaatc tggaataatg gtcatccttt ttctgaacaa gcaaaaaaaa atgctggaaa
     2341 actatttttc acagaagata ctggacgtag tgggaaacac tatgggattg gactatcttt
     2401 tgctcaaggt gtagctttaa aacatcaagg aaacttaatt ctcagtaatc ctcaaaaagg
     2461 tggggcagaa gttatcctaa aaataaaaaa gtaatttagt aatctctaag gattactttt
     2521 tttgtttcgg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2581 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2641 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2701 caccggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2761 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2821 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2881 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2941 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3001 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3061 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3121 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3181 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3241 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3301 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3361 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3421 aggcgggcag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3481 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3541 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3601 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3661 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3721 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3781 aaccgcctct ccccgcgcgt tggccgattc attaatgcag gttaacctgg cttatcgaaa
     3841 ttaatacgac tcactatagg gagaccggcc tcgagaagct cgcgttatcg gtcctttaat
     3901 tggtggacaa atttacatta gtctcggaca ttctgctccc gcccttatgg gatttatctt
     3961 ccttattctc gctttgattg ttctatcgaa agcgaaatca aacgaataat ttataaataa
     4021 aaaaaccacc tctaaaaggt gatttttatt tataaattac aatttttcag caatatcagt
     4081 aattgcttta tcaactgctg ctttttggct atcaatcaaa gcaacacgtg ctgtaatttg
     4141 tttaattgcc atttcaattg aacgtttcaa gccttggttt tctaattttg gttcaaagaa
     4201 agcttgagct ctctagaact agtggtaccg catgcctgca gatccggtct aacaaagaaa
     4261 aacacatttt tttgtgaaaa ttcgttttta ttattcaaca tagttccctt caagagcgat
     4321 acaacgatta taacgacctt ccaatttttt gataccattt tggtagtact ccttcggttt
     4381 tgcctcaaaa taggcctcag tttcggcgat cacctcttca ttgcagccaa attttttccc
     4441 tgcgagcatc cttttgaggt ctgagaacaa gaaaaagtcg ctgggggcca gatctggaga
     4501 atacggcggg tggggaagca attcgaagcc caattcatga atttttgcca tcgttctcaa
     4561 tgacttgtgg cacggtgcgt tgtcttggtg gaacaacact tttttcttct tcatgtgggg
     4621 ccgttttgcc gcgatttcga ccttcaaacg ctccaataac gccatataat agtcactgtt
     4681 gatggttttt cccttctcaa gataatcgat aaaaattatt ccatgcgcat cccaaaaaac
     4741 agaggccatt actttgccag cggacttttg agtctttcca cgcttcggag acggttcacc
     4801 ggtcgctgtc cactcagccg actgtcgatt ggactcagga gtgtagtgat ggagccatgt
     4861 ttcatccatt gtcacatatc gacggaaaaa ctcgggtgta ttacgagtta acagctgcaa
     4921 acaccgctta gaatcatcaa cacgtcgttg tttttggtca aatgtgagct cgcgcggcac
     4981 ccatttcgca cagagcttcc gcatatccaa atattgatga atgatatgac caacacgttc
     5041 ctttgatatc tttaaggcct ctgctatctc gatcaacttc attttacggt cattcaaaat
     5101 cattttgtgg atttttttga tgttttcgtc ggtaaccacc tctttcgggc gtccactgcg
     5161 ttcaccgtcc tccgtgctca tttcaccacg cttgaatttt gcataccaat caattattgt
     5221 tgatttccct ggggcagagt ccggaaactc attatcaagc caagtttttg cttccactgt
     5281 attttttccc ttcagaaaac agtattttat caaaacacga aattcctttt tttccatggt
     5341 gagtgcctcc ttataattta ttttgtagtt ccttcgaacg aaatcattgt atctaacaaa
     5401 cttcagaatt taatcagagc cgtttattat gctcgcgtta tcgacaataa tattattacc
     5461 aatactttct caagatagaa ttaagactgt tttagatttg ttaatgtttc tattgtcagt
     5521 atagttataa gactagatct gatccgtagc ggttttcgat tcacgggaat tcgacttctc
     5581 tactcctttt aatcaaaaat ctgattgact gtaaataaga atttagtaat tatagttaat
     5641 tcatttttgt ttcatgctat ttccatataa aaatcttgca taacctatct ccgcggtaac
     5701 aggttggatg ataagtcccc ggtctgacac atagatggcg tcgctagtat taaatgcata
     5761 ttatttttat atagtaccaa ccttcaaatg attcccgccc tcctatagtg agtcgtatta
     5821 aaacccagcg aaccatttga ggtgataggt aagattatac cgaggtatga aaacgagaat
     5881 tggaccttta cagaattact ctatgaagcg ccatatttaa aaagctacca agacgaagag
     5941 gatgaagagg atgaggaggc agattgcctt gaatatattg acaatactga taagataata
     6001 tatcttttat atagaagata tcgccgtatg taaggatttc agggggcaag gcataggcag
     6061 cgcggggcga gctcggtacc cggggatcct ctagagtcaa ttccccttgc catgtatatg
     6121 aaaaattagg tggtaaaaat gattataaag atgaaatagt atatgtatat gattatgaaa
     6181 aaggtgataa ataaatgaat atagttgaaa atgaaatatg tataagaact ttaatagatg
     6241 atgattttcc tttgatgtta aaatggttaa ctgatgaaag agtattagaa ttttatggtg
     6301 gtagagataa aaaatataca ttagaatcat taaaaaaaca ttatacagag ccttgggaag
     6361 atgaagtttt tagagtaatt attgaatata acaatgttcc tattggatat ggacaaatat
     6421 ataaaatgta tgatgagtta tatactgatt atcattatcc aaaaactgat gagatagtct
     6481 atggtatgga tcaatttata ggagagccga attattggag taaaggaatt ggtacaagat
     6541 atattaaatt gatttctgaa tttttgaaaa aagaaagaaa tgctaatgca gttattttag
     6601 accctcataa aaataatcca agagcaataa gggcatacca aagatctggt tttagaatta
     6661 ttgaagattt gccagaacat gaattacacg agggcaaaaa agaagattgt tatttaatgg
     6721 aatatagata tgatgataat gccacaaatg ttaaggcaat gaaatattta attgagcatt
     6781 actttgataa tttcaaagta gatagtattg aaataatcgg tagtggttat gatagtgtgg
     6841 catatttagt taataatgaa tacattttta aaacaaaatt tagtactaat aagaaaaaag
     6901 gttatgcaaa agaaaaagca atatataatt ttttaaatac aaatttagaa actaatgtaa
     6961 aaattcctaa tattgaatat tcgtatatta gtgatgaatt atctatacta ggttataaag
     7021 aaattaaagg aactttttta acaccagaaa tttattctac tatgtcagaa gaagaacaaa
     7081 atttgttaaa acgagatatt gccagttttt taagacaaat gcacggttta gattatacag
     7141 atattagtga atgtactatt gataataaac aaaatgtatt agaagagtat atattgttgc
     7201 gtgaaactat ttataatgat ttaactgata tagaaaaaga ttatatagaa agttttatgg
     7261 aaagactaaa tgcaacaaca gtttttgagg gtaaaaagtg tttatgccat aatgatttta
     7321 gttgtaatca tctattgtta gatggcaata atagattaac tgggataatt gattttggag
     7381 attctggaat tatagatgaa tattgtgatt ttatatactt acttgaagat agtgaagaag
     7441 aaataggaac aaattttgga gaagatatat taagaatgta tggaaatata gatattgaga
     7501 aagcaaaaga atatcaagat atagttgaag aatattatcc tattgaaact attgtttatg
     7561 gaattaaaaa tattaaacag gaatttatcg aaaatggtag aaaagaaatt tataaaagga
     7621 cttataaaga ttgactatat aatatatgaa aagggggatc gacctgcagg catgcaagct
     7681 tggcgtaatc atggtcatag ctgtttcctg taatacgact cactatagga gggcgggtat
     7741 agatctaaaa cgaattttaa caaaaaaatg tgtttttctt tgttagaccg gggacttatc
     7801 atccaacctg ttaccgcgga tgctctagag gatcattttg tttattgcaa ttgtattgct
     7861 attaatcgca acatcaaacc aaaataaaaa cccccttcga ctttcttaag ctggcgtaat
     7921 agcgaagagg cccgcaccgg gggatcctct agagtcgacc tgcaggtcga taaacccagc
     7981 gaaccatttg aggtgatagg taagattata ccgaggtatg aaaacgagaa ttggaccttt
     8041 acagaattac tctatgaagc gccatattta aaaagctacc aagacgaaga ggatgaagag
     8101 gatgaggagg cagattgcct tgaatatatt gacaatactg ataagataat atatctttta
     8161 tatagaagat agcttgatcg gcaatagtta cccttattat caagataaga aagaaaagga
     8221 tttttcgcta cgctcaaatc ctttaaaaaa acacaaaaga ccacattttt taatgtggtc
     8281 ttttattctt caactaaagc acccattagt tcaacaaacg aaaattggat aaagtgggat
     8341 atttttaaaa tatatattta tgttacagta atattgactt ttaaaaaagg attgattcta
     8401 atgaagaaag cagacaagta agcctcctaa attcacttta gataaaaatt taggaggcat
     8461 atcaaatgaa ctttaataaa attgatttag acaattggaa gagaaaagag atatttaatc
     8521 attatttgaa ccaacaaacg acttttagta taaccacaga aattgatatt agtgttttat
     8581 accgaaacat aaaacaagaa ggatataaat tttaccctgc atttattttc ttagtgacaa
     8641 gggtgataaa ctcaaataca gcttttagaa ctggttacaa tagcgacgga gagttaggtt
     8701 attgggataa gttagagcca ctttatacaa tttttgatgg tgtatctaaa acattctctg
     8761 gtatttggac tcctgtaaag aatgacttca aagagtttta tgatttatac ctttctgatg
     8821 tagagaaata taatggttcg gggaaattgt ttcccaaaac acctatacct gaaaatgctt
     8881 tttctctttc tattattcca tggacttcat ttactgggtt taacttaaat atcaataata
     8941 atagtaatta ccttctaccc attattacag caggaaaatt cattaataaa ggtaattcaa
     9001 tatatttacc gctatcttta caggtacatc attctgtttg tgatggttat catgcaggat
     9061 tgtttatgaa ctctattcag gaattgtcag ataggcctaa tgactggctt ttataatatg
     9121 agataatgcc gactgtactt tttacagtcg gttttctaat gtcactaacc tgccccgtta
     9181 gttgaagaag gtttttatat tacagctcca gatcatcgaa ttatcgggga agaacagtat
     9241 gtcgagctat tttttgactt actggggatc aagcctgatt gggagaaaat aaaatattat
     9301 attttactgg atgaattgtt ttagtaccta gatttagatg tctaaaaagc tttaactaca
     9361 agctttttag acatctaatc ttttctgaag tacatccgca actgtccata ctctgatgtt
     9421 ttatatcttt tctaaaagtt cgctagatag gggtcccgag cgcctacgag gaatttgtat
     9481 cgactctaga ggatcatttt gtttattgca attgtattgc tattaatcgc aacatcaaac
     9541 caaaataaaa acccccttcg actttcgtca gggggctttt atttattcaa taatccctcc
     9601 tctcaataaa tctattgttg tacttaattc aacttccatt tctctgtatc tttcaatacg
     9661 ctcttttaag tccttaattt ctttttttaa ttcctcattt tcagcaaata actctttttc
     9721 tttgtttgtc attttatttc ccccgtttca gcatcaagaa cctttgcata acttgctcta
     9781 tatccacact gataattgcc ctcaaaccat aatctaaagg cgctagagtt tgttgaaaca
     9841 atatctttta catcattcgt atttaaaatt ccaaactccg ctcccctaag gcgaataaaa
     9901 gccattaaat cttttgtatt taccaaatta tagtcatcca ctatatctaa aagtaaattc
     9961 ttcaattctc ttttttggct ttcatcaagt gttatatagc ggtcaatatc aaaatcatta
    10021 atgttcaaaa tatctttttt gtcgtatata tgtttattct tagcaatagc gtcctttgat
    10081 tcatgagtca aatattcata tgaacctttg atataatcaa gtatctcaac atgagcaact
    10141 gaactattcc ccaattttcg cttaatcttg ttcctaacgc tttctattgt tacaggattt
    10201 cgtgcaatat atataacgtg atagtgtggt tttttatagt gctttccatt ttgtataata
    10261 ttactattat tccatgtatc tttatctttt ttttcgtcca tatcgtgtaa aggactgaca
    10321 gccatagata cgcccaaact ctctaatttt tctttccaat cattaggaat tgagtcagga
    10381 tataataaaa atccaaaatt tctagcttta gtatttttaa tagccatgat ataattacct
    10441 tatcaaaaac aagtagcgaa aactcgtatc cttctaaaaa cgcgagcttt cgcttatttt
    10501 ttttattctg attcctttct tgcatattct tctatagcta acgccgcaac cgcagatttt
    10561 gaaaaacctt tttgtttcgc catatctgtt aattttttat cttgctcttt tgtcagagaa
    10621 atcataactc tttttttcga ttctgaaatc accatttaaa aaactccaat caaataattt
    10681 tataaagtta gtgtatcact ttgtaatcat aaaaacaaca ataaagctac ttaaatatag
    10741 atttataaaa aacgttggcg aaaacgttgg cgattcgttg gcgattgaaa aaccccttaa
    10801 acccttgagc cagttgggat agagcgtttt tggcacaaaa attggcactc ggcacttaat
    10861 ggggggtcgt agtacggaag caaaattcgc ttcctttccc cccatttttt tccaaattcc
    10921 aaattttttt caaaaatttt ccagcgctac cgctcggcaa aattgcaagc aatttttaaa
    10981 atcaaaccca tgagggaatt tcattccctc atactccctt gagcctcctc caaccgaaat
    11041 agaagggcgc tgcgcttatt atttcattca gtcatcggct ttcataatct aacagacaac
    11101 atcttcgctg caaagccacg ctacgctcaa gggcttttac gctacgataa cgcctgtttt
    11161 aacgattatg ccgataacta aacgaaataa acgctaaaac gtctcagaaa cgattttgag
    11221 acgttttaat aaaaaatcgg ataataaata tatataaacg tatatagatt tcataaagtc
    11281 taacacacta gacttattta cttcgtaatt aagtcgttaa accgtgtgct ctacgaccaa
    11341 aactataaaa cctttaagaa ctttcttttt ttacaagaaa aaagaaatta gataaatctc
    11401 tcatatcttt tattcaataa tcgcatccga ttgcagtata aatttaacga tcccccggag
    11461 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
    11521 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
    11581 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
    11641 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
    11701 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
//
