LOCUS       pInvRecA.        7354 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid for transiently converting any E. coli strain to a RecA+ 
            phenotype for phage P1-mediated transduction.
ACCESSION   .
VERSION     .
KEYWORDS    pInvRecA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7354)
  AUTHORS   Sektas M, Gregorowicz M, Szybalski W.
  TITLE     Transient conversion to RecA+ phenotype to permit P1 transduction in
            any Escherichia coli recA- strains.
  JOURNAL   BioTechniques 1999;27:911-4.
  PUBMED    10572635
REFERENCE   2  (bases 1 to 7354)
  AUTHORS   Lucigen
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7354)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "1999"; volume: "27"; pages: "911-4"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     Addition of heated chlortetracycline induces synthesis of Flp 
            recombinase, thereby inverting the recA gene so that it is 
            expressed.
            Growth at 42(o)C inactivates the temperature-sensitive mutant Rep101
            protein, causing loss of the plasmid.
FEATURES             Location/Qualifiers
     source          1..7354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(72..695)
                     /label=TetR
                     /note="tetracycline repressor TetR"
     promoter        711..766
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             783..2048
                     /label=FLP
                     /note="site-specific recombinase"
     protein_bind    2251..2284
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             complement(2314..3372)
                     /label=RecA
                     /note="DNA repair protein from E. coli"
     RBS             complement(3379..3401)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(3425..3458)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     terminator      3602..3645
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3743..3829
                     /gene="Escherichia coli rrnB"
                     /label=Escherichia coli rrnB terminator
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3924..4010
                     /gene="Escherichia coli rrnB"
                     /label=Escherichia coli rrnB terminator
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(4105..4191)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(4519..5466)
                     /label=Rep101(Ts)
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
     rep_origin      complement(5514..5736)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(6361..7218)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7219..7323)
                     /label=AmpR promoter
ORIGIN
        1 cgatgggtgg ttaactcgac atcttggtta ccgtgaagtt accatcacgg aaaaaggtta
       61 tgctgctttt aagacccact ttcacattta agttgttttt ctaatccgca tatgatcaat
      121 tcaaggccga ataagaaggc tggctctgca ccttggtgat caaataattc gatagcttgt
      181 cgtaataatg gcggcatact atcagtagta ggtgtttccc tttcttcttt agcgacttga
      241 tgctcttgat cttccaatac gcaacctaaa gtaaaatgcc ccacagcgct gagtgcatat
      301 aatgcattct ctagtgaaaa accttgttgg cataaaaagg ctaattgatt ttcgagagtt
      361 tcatactgtt tttctgtagg ccgtgtacct aaatgtactt ttgctccatc gcgatgactt
      421 agtaaagcac atctaaaact tttagcgtta ttacgtaaaa aatcttgcca gctttcccct
      481 tctaaagggc aaaagtgagt atggtgccta tctaacatct caatggctaa ggcgtcgagc
      541 aaagcccgct tattttttac atgccaatac aatgtaggct gctctacacc tagcttctgg
      601 gcgagtttac gggttgttaa accttcgatt ccgacctcat taagcagctc taatgcgctg
      661 ttaatcactt tacttttatc taatctagac atcattaatt cctaattttt gttgacactc
      721 tatcattgat agagttattt taccactccc tatcagtgat agagaaaagt gaaatgaatt
      781 cgccacaatt tggtatatta tgtaaaacac cacctaaggt gcttgttcgt cagtttgtgg
      841 aaaggtttga aagaccttca ggtgagaaaa tagcattatg tgctgctgaa ctaacctatt
      901 tatgttggat gattacacat aacggaacag caatcaagag agccacattc atgagctata
      961 atactatcat aagcaattcg ctgagtttcg atattgtcaa taaatcactc cagtttaaat
     1021 acaagacgca aaaagcaaca attctggaag cctcattaaa gaaattgatt cctgcttggg
     1081 aatttacaat tattccttac tatggacaaa aacatcaatc tgatatcact gatattgtaa
     1141 gtagtttgca attacagttc gaatcatcgg aagaagcaga taagggaaat agccacagta
     1201 aaaaaatgct taaagcactt ctaagtgagg gtgaaagcat ctgggagatc actgagaaaa
     1261 tactaaattc gtttgagtat acttcgagat ttacaaaaac aaaaacttta taccaattcc
     1321 tcttcctagc tactttcatc aattgtggaa gattcagcga tattaagaac gttgatccga
     1381 aatcatttaa attagtccaa aataagtatc tgggagtaat aatccagtgt ttagtgacag
     1441 agacaaagac aagcgttagt aggcacatat acttctttag cgcaaggggt aggatcgatc
     1501 cacttgtata tttggatgaa tttttgagga attctgaacc agtcctaaaa cgagtaaata
     1561 ggaccggcaa ttcttcaagc aataaacagg aataccaatt attaaaagat aacttagtca
     1621 gatcgtacaa taaagctttg aagaaaaatg cgccttattc aatctttgct ataaaaaatg
     1681 gcccaaaatc tcacattgga agacatttga tgacctcatt tctttcaatg aagggcctaa
     1741 cggagttgac taatgttgtg ggaaattgga gcgataagcg tgcttctgcc gtggccagga
     1801 caacgtatac tcatcagata acagcaatac ctgatcacta cttcgcacta gtttctcggt
     1861 actatgcata tgatccaata tcaaaggaaa tgatagcatt gaaggatgag actaatccaa
     1921 ttgaggagtg gcagcatata gaacagctaa agggtagtgc tgaaggaagc atacgatacc
     1981 ccgcatggaa tgggataata tcacaggagg tactagacta cctttcatcc tacataaata
     2041 gacgcatata agtacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat
     2101 atatatacag gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcatcg
     2161 tccattccga cagcatcgcc agtcactatg gcgtgctgct agcatcgatc catggactag
     2221 tctcgagttt aaagatcgat gaattgatcc gaagttccta ttctctagaa agtataggaa
     2281 cttcgaattg tcgagtgcgg ccgcaagctt ttaaaaatct tcgttagttt ctgctacgcc
     2341 ttcgctatca tctacagaga aatccggcgt tgagttcggg ttgctcagca gcaactcacg
     2401 tactttcttc tcgatctctt tcgcggtttc cgggttatct ttcagccagg cagtcgcatt
     2461 cgctttaccc tgaccgatct tctcaccttt gtagctgtac cacgcgcctg ctttctcgat
     2521 cagcttctct tttacgccca ggtcaaccag ttcgccgtag aagttgatac cttcgccgta
     2581 gaggatctgg aattcagcct gtttaaacgg cgcagcgatt ttgttcttca ccactttcac
     2641 gcgggtttcg ctacccacca cgttttcgcc ctctttcacc gcgccgatac gacggatgtc
     2701 gagacgaaca gaggcgtaga atttcagcgc gttaccaccg gtagtggttt ccgggttacc
     2761 gaacatcaca ccaattttca tacggatctg gttgatgaag atcagcagcg tgttggactg
     2821 cttcaggtta cccgccagct tacgcatcgc ctggctcatc atacgtgccg caaggcccat
     2881 gtgagagtcg ccgatttcgc cttcgatttc cgctttcggc gtcagtgccg ccacggagtc
     2941 aacgacgata acgtctactg cgccagaacg cgccagggcg tcacagattt ccagtgcctg
     3001 ctcgccggtg tccggctggg agcacagcag gttgtcgata tcgacgccca gtttacgtgc
     3061 gtagattggg tccagcgcgt gttcagcatc gataaacgca caggttttac cttcacgctg
     3121 cgctgcggcg atcacctgca gcgtcagcgt ggttttaccg gaagattccg gtccgtagat
     3181 ctcgacgata cggcccatcg gcagaccacc tgccccaagc gcgatatcca gtgaaagcga
     3241 accggtagag atggtttcca catccattga acggtcttca cccaggcgca tgatggagcc
     3301 tttaccaaat tgtttctcaa tctggcccag tgctgccgcc aacgctttct gtttgttttc
     3361 gtcgatagcc atggtatatc tccttcttaa agttaaacaa aattatttct aggatcccca
     3421 attcgaagtt cctatacttt ctagagaata ggaacttcgg atcgggaatt ccgatcccca
     3481 attcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcgcaac
     3541 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     3601 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggaatt
     3661 aattcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa
     3721 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     3781 agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggaat
     3841 taattcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcgca
     3901 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
     3961 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggaa
     4021 ttaattcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc
     4081 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     4141 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctgga
     4201 attgggaatt aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4261 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4321 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4381 accctgataa atgcttcaat aatctagccc atgggtatgg acagttttcc ctttgatatg
     4441 taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact gatagataca
     4501 agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta gtgtggttcg
     4561 ttgtttttgc gtgagccatg agaacgaacc attgagatca tgcttacttt gcatgtcact
     4621 caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt
     4681 ttttcttagt ccgttacgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc
     4741 attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc
     4801 aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt
     4861 gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa
     4921 ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat
     4981 atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat
     5041 agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa
     5101 ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa
     5161 cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc
     5221 cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct
     5281 actgatgttc atcatctgaa cgtattggtt ataagtgaac gataccgtcc gttctttcct
     5341 tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc
     5401 atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc
     5461 agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag
     5521 tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat tctgctagac
     5581 ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg
     5641 tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa
     5701 gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta
     5761 ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa
     5821 aggcttaagt agcaccctcg caagctcggt tgcggccgca atcgggcaaa tcgctgaata
     5881 ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc
     5941 gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg
     6001 attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt
     6061 tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc
     6121 tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa
     6181 tgcacccagt aaggcagcgg tatcatcaac ggggtctgac gctcagtgga acgaaaactc
     6241 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6301 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6361 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6421 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6481 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6541 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6601 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6661 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6721 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6781 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6841 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6901 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6961 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7021 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7081 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7141 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7201 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7261 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7321 gcgcacattt ccccgaaaag tgccacttgc atcg
//
