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" 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" primer_bind complement(8240..8259) /label=T7 /note="T7 promoter, forward primer" 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 //