LOCUS       Exported                6555 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  tetracycline-inducible expression vector for Staphylococcus aureus.
ACCESSION   .
VERSION     .
KEYWORDS    pRMC2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6555)
  AUTHORS   Corrigan RM, Foster TJ
  TITLE     An improved tetracycline-inducible expression vector for 
            Staphylococcus aureus.
  JOURNAL   Plasmid. 2009 Mar;61(2):126-9. doi: 10.1016/j.plasmid.2008.10.001. 
            Epub 2008 Nov 25.
  PUBMED    18996145
REFERENCE   2  (bases 1 to 6555)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6555
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(18..40)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     140..159
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     368..384
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     495..511
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    562..580
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             690..1313
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(763..782)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             complement(1625..1639)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(4323..4339)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4323..4339)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4336..4358)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4347..4363
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4371..4401)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4416..4437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4554..4571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4725..5313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4805..4824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5484..6344)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6107..6126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6345..6449)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6517..6535
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
       61 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
      121 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
      181 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
      241 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      301 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      361 cgacgttgta aaacgacggc cagtgaattc gagctcagat ctgttaacgg taccatcaag
      421 cttattttaa ttatactcat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      481 ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcagatctg ttaacggtac
      541 catcaagctt attttaatta tactctatca atgatagagt gtcaatattt tttttagttt
      601 ttcatgaact cgaggggatc caaataaaaa actagtttga caaataactc tatcaatgat
      661 ataatgtcaa caaaaaggag gaattaatga tgtctagatt agataaaagt aaagtgatta
      721 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
      781 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
      841 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
      901 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
      961 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     1021 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     1081 tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     1141 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     1201 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     1261 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtct taaaagcagc
     1321 ataacctttt tccgtgatgg taacttcacg gtaaccaaga tgtcgagtta tcgataccgt
     1381 cgacctgcag gcatgcaagc ttttaaaaag caaatatgag ccaaataaat atattctaat
     1441 tctacaaaca aaaatttgag caaattcagt gtcgattttt taagacactg cccagttaca
     1501 tgcaaattaa aattttcatg attttttata gttcctaaca gggttaaaat ttgtataacg
     1561 aaagtataat gtttatataa cgttagtata ataaagcatt ttaacattat acttttgata
     1621 atcgtttatc gtcgtcatca caataacttt taaaatactc gtgcataatt caacagctga
     1681 cctcccaata actacatggt gttatcggga ggtcagctgt tagcacttat attttgttat
     1741 tgttcttcct cgatttcgtc tatcattttg tgattaattt ctcttttttc ttgttctgtt
     1801 aagtcataaa gttcactagc taaatactct ttttgtttcc aaatataaaa aatttgatag
     1861 atatattcgg ttggatcaat ttcttttaag taatctaaat ccccattttt taatttcttt
     1921 ttagcctctt taaataatcc tgaataaact aatacctgtt tacctttaag tgatttataa
     1981 aatgcatcaa agactttttg atttattaaa taatcactat ctttaccaga atacttagcc
     2041 atttcatata attctttatt attattttgt cttatttttt gaacttgaac ttgtgttatt
     2101 tctgaaatgc ccgttacatc acgccataaa tctaaccatt cttgttggct aatataatat
     2161 cttttatctg tgaaatacga tttatttact gcaattaaca catgaaaatg aggattataa
     2221 tcatctcttt ttttattata tgtaatctct aacttacgaa catatccctt tataacacta
     2281 cctacttttt ttctctttat aagttttcta aaagaattat tataacgttt tatttcattt
     2341 tctaattcat cactcattac attaggtgta gtcaaagtta aaaagataaa ctcctttttc
     2401 tcttgctgct taatatattg catcatcaaa gataaaccca atgcatcttt tctagctttt
     2461 ctccaagcac agacaggaca aaatcgattt ttacaagaat tagctttata taatttctgt
     2521 ttttctaaag ttttatcagc tacaaaagac agaaatgtat tgcaatcttc aactaaatcc
     2581 atttgattct ctccaatatg acgtttaata aatttctgaa atacttgatt tctttgtttt
     2641 ttctcagtat acttttccat gttataacac ataaaaacaa cttagttttc acaaactatg
     2701 acaataaaaa aagttgcttt ttcccctttc tatgtatgtt ttttactatt catttaaaac
     2761 gatacattaa taggtacgaa aaagcaactt tttttgcgct taaaaccagt cataccaata
     2821 acttaagggt aactagcctc gccggcaata gttaccctta ttatcaagat aagaaagaaa
     2881 aggatttttc gctacgctca aatcctttaa aaaaacacaa aagaccacat tttttaatgt
     2941 ggtcttttat tcttcaacta aagcacccat tagttcaaca aacgaaaatt ggataaagtg
     3001 ggatattttt aaaatatata tttatgttac agtaatattg acttttaaaa aaggattgat
     3061 tctaatgaag aaagcagaca agtaagcctc ctaaattcac tttagataaa aatttaggag
     3121 gcatatcaaa tgaactttaa taaaattgat ttagacaatt ggaagagaaa agagatattt
     3181 aatcattatt tgaaccaaca aacgactttt agtataacca cagaaattga tattagtgtt
     3241 ttataccgaa acataaaaca agaaggatat aaattttacc ctgcatttat tttcttagtg
     3301 acaagggtga taaactcaaa tacagctttt agaactggtt acaatagcga cggagagtta
     3361 ggttattggg ataagttaga gccactttat acaatttttg atggtgtatc taaaacattc
     3421 tctggtattt ggactcctgt aaagaatgac ttcaaagagt tttatgattt atacctttct
     3481 gatgtagaga aatataatgg ttcggggaaa ttgtttccca aaacacctat acctgaaaat
     3541 gctttttctc tttctattat tccatggact tcatttactg ggtttaactt aaatatcaat
     3601 aataatagta attaccttct acccattatt acagcaggaa aattcattaa taaaggtaat
     3661 tcaatatatt taccgctatc tttacaggta catcattctg tttgtgatgg ttatcatgca
     3721 ggattgttta tgaactctat tcaggaattg tcagataggc ctaatgactg gcttttataa
     3781 tatgagataa tgccgactgt actttttaca gtcggttttc taatgtcact aacctgcccc
     3841 gttagttgaa gaaggttttt atattacagc tccagatcca tatccttctt tttctgaacc
     3901 gacttctcct ttttcgcttc tttattccaa ttgctttatt gacgttgagc ctcggaaccc
     3961 ttaacaatcc caaaacttgt cgaatggtcg gcttaatagc tcacgctatg ccgacattcg
     4021 tctgcaagtt tagttaaggg ttcttctcaa cgcacaataa attttctcgg cataaatgcg
     4081 tggtctaatt tttattttta ataaccttga tagcaaaaaa tgccattcca atacaaaacc
     4141 acatacctat aatcgataac cacataacag tcataaaacc actccttttt aacaaacttt
     4201 atcacaagaa atatttaaat tttaaatgcc tttattttga attttaaggg gcattttaaa
     4261 gatttagggg taaatcatat agttttatgc ctaaaaacct acagaagctt ggcgtaatca
     4321 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4381 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4441 gcgttgcgct cactgtccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4501 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4561 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4621 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4681 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4741 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4801 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4861 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4921 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4981 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5041 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5101 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5161 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5221 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5281 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5341 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5401 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5461 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5521 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5581 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5641 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5701 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5761 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5821 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5881 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5941 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6001 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6061 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6121 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6181 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6241 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6301 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6361 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6421 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6481 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6541 cgtctcgcgc gtttc
//
