LOCUS       pLPT234                 9745 bp    DNA     circular UNA 21-MAY-2019
DEFINITION  repressilatore without degradations tags, with triple reporter.
ACCESSION   urn.local...m-9uq5zle
VERSION     urn.local...m-9uq5zle
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     rep_origin      complement(9743..220)
     terminator      complement(712..806)
     CDS             complement(836..1696)
                     /standard_name="AmpR"
     CDS             complement(1738..1766)
                     /standard_name="pAmpR"
     terminator      complement(1940..2026)
     CDS             complement(2047..2769)
                     /standard_name="mSCFP3"
     promoter        complement(2823..2868)
     regulatory      complement(2823..2828)
                     /standard_name="-10"
     regulatory      complement(2829..2845)
                     /standard_name="LacO"
     regulatory      complement(2846..2851)
                     /standard_name="-35"
     regulatory      complement(2852..2868)
                     /standard_name="LacO"
     misc_feature    2969..3013
                     /standard_name="Buffer from R1"
     promoter        3025..3073
     regulatory      3025..3040
                     /standard_name="OR2"
     regulatory      3038..3043
                     /standard_name="-35"
     regulatory      3048..3063
                     /standard_name="OR1"
     regulatory      3059..3064
                     /standard_name="-10"
     misc_feature    3086..3134
                     /standard_name="5'UTR and RBS"
     misc_feature    3135..3167
                     /standard_name="n-term from mCherry"
     CDS             3135..3857
                     /standard_name="mKate2 hybrid (mCherry N-term)"
     terminator      3891..3923
     terminator      complement(3971..4057)
     CDS             complement(4118..4834)
                     /standard_name="mVenus"
     RBS             complement(4843..4854)
     promoter        complement(4864..4937)
     protein_bind    complement(4894..4912)
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /standard_name="tet operator"
     protein_bind    complement(4919..4937)
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /standard_name="tet operator"
     protein_bind    complement(5055..5071)
                     /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)."
                     /standard_name="lac operator"
     promoter        5055..5100
     regulatory      5072..5077
                     /standard_name="-35"
     protein_bind    complement(5078..5094)
                     /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)."
                     /standard_name="lac operator"
     regulatory      5095..5100
                     /standard_name="-10"
     RBS             5136..5147
     CDS             5154..5777
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by
                     adding tetracycline or doxycycline."
                     /standard_name="TetR"
     terminator      complement(5804..5890)
     promoter        5931..5979
     regulatory      5931..5946
                     /standard_name="OR2"
     regulatory      5944..5949
                     /standard_name="-35"
     regulatory      5954..5969
                     /standard_name="OR1"
     regulatory      5965..5970
                     /standard_name="-10"
     RBS             5997..6008
     CDS             6026..7108
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /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)."
                     /standard_name="lacI"
     terminator      complement(7135..7221)
     promoter        7252..7325
     protein_bind    complement(7277..7295)
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /standard_name="tet operator"
     RBS             7335..7346
     CDS             7353..8066
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /standard_name="-lambda repressor"
     terminator      complement(8124..8210)
     CDS             complement(8745..9695)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /standard_name="Rep101"
ORIGIN      
        1 ctagtcaatg ataattacta gtccttttcc tttgagttgt gggtatctgt aaattctgct
       61 agacctttgc tggaaaactt gtaaattctg ctagaccctc tgtaaattcc gctagacctt
      121 tgtgtgtttt ttttgtttat attcaagtgg ttataattta tagaataaag aaagaataaa
      181 aaaagataaa aagaatagat cccagccctg tgtataactc actactttag tcagttccgc
      241 agtattacaa aaggatgtcg caaacgctgt ttgctcctct acaaaacaga ccttaaaacc
      301 ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct
      361 ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg
      421 gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat tcatgcaagg
      481 aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt tatggcgggt
      541 ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca
      601 gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa
      661 ggcagcggta tcatcaacag gcttacccgt cttactgtcc ctagtgcttg gattctcacc
      721 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
      781 cattactgga tctatcaaca ggagtccaag cgagctcgta aacttggtct gacagttacc
      841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     1741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1801 gcacatttcc ccgaaaagtg ccacctgacg tctctttcct gcgttatccc ctgattctgt
     1861 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc cagcctacac
     1921 cccgggtcta gggcggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     1981 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctctagactc
     2041 agctaattaa gcttatttgt atagttcatc catgccatgt gtaatcccag cagctgttac
     2101 aaactcaaga aggaccatgt ggtctctctt ttcgttggga tctttcgaaa gcttagattg
     2161 tgtggacagg taatggttgt ctggtaaaag gacagggcca tcgccaattg gagtattttg
     2221 ttgataatgg tctgctagtt gaacgcctcc atcttcaatg ttgtgtctaa ttttgaagtt
     2281 agctttgatt ccattctttt gtttgtctgc cgtgatgtat acattgtctg agatgtagtt
     2341 gtattccaat ttgtgtccaa gaatgtttcc atcttcttta aaatcaatac cttttaactc
     2401 gattctatta acaagggtat caccttcaaa cttgacttca gcacgtgtct tgtagttccc
     2461 gtcatctttg aaaaatatag ttctttcctg tacataacct tcgggcatgg cactcttgaa
     2521 aaagtcatgc tgtttcatat gatctgggta tcttgcaaag cattgaacac cccaagtgag
     2581 agtagtgaca agtgttggcc atggaacagg tagttttcca gtagtgcaaa taaatttaag
     2641 ggtaagtttt ccgtatgttg catcaccttc accctctcca ctgacagaaa atttgtgccc
     2701 attaacatca ccatctaatt caacaagaat tgggacaact ccagtgaaaa gttcttctcc
     2761 tttactcatg gtacctttct cctctttaat gaattcggtc agtgcgtcct gctgatgtgc
     2821 tcagtatctt gttatccgct cacaatgtca attgttatcc gctcacaatt ctcgaggtga
     2881 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     2941 tcttagacgt caggtggcac ttttcgggtc acaaaagaca ttccaccaaa cgtcgtggcg
     3001 gctggcgttc cttggataaa tatctaacac cgtgcgtgtt gactatttta cctctggcgg
     3061 tgataatggt tgcatgtact agaatatcag caggacgcac tgaccgaatt cattaaagag
     3121 gagaaaggta ccgcatggtt agtaaaggag aagaaaataa catggcactg attaaggaga
     3181 acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat
     3241 ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag gccgtcgagg
     3301 gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa
     3361 ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc cctgagggct
     3421 tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
     3481 ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat
     3541 ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggcctccacc gagaccctgt
     3601 accccgctga cggcggcctg gaaggcagag ccgacatggc cctgaagctc gtgggcgggg
     3661 gccacctgat ctgcaacttg aagaccacat acagatccaa gaaacccgct aagaacctca
     3721 agatgcccgg cgtctactat gtggacagaa gactggaaag aatcaaggag gccgacaaag
     3781 agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc cctagcaaac
     3841 tggggcacag agtctagggg atcccccggg ctgcaggaat tcgatatcac gcaaaaaacc
     3901 ccgcttcggc ggggtttttt cgcgagctcg aattcactgg ccgtcgtttt acataggtct
     3961 agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     4021 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctctagcac gcgtaccatg
     4081 ggatcccccg ggctgcagga attcgatatc aagcttatta tttgtatagt tcatccatgc
     4141 catgtgtaat cccagcagct gttacaaact caagaaggac catgtggtct ctcttttcgt
     4201 tgggatcttt cgaaagctta gattggtagg acaggtaatg gttgtctggt aaaaggacag
     4261 ggccatcgcc aattggagta ttttgttgat aatggtctgc tagttgaaca cctccatctt
     4321 caatgttgtg tctaattttg aagttcgctt tgattccatt cttttgtttg tctgccgtga
     4381 tgtatacatt gtgtgagtta tagttgtatt ccaatttgtg tccaagaatg tttccatctt
     4441 ctttaaaatc aatacctttt aactcgattc tattaacaag ggtatcacct tcaaacttga
     4501 cttcagcacg tgtcttgtag ttcccgtcat ctttgaaaaa tatagttctt tcctgtacat
     4561 aaccttcggg catggcactc ttgaaaaagt catgctgttt catatgatct gggtatctcg
     4621 caaagcattg aacaccataa cccaaagtag tgacaagtgt tggccatgga acaggtagtt
     4681 ttccagtagt gcaaatcaat ttaagggtaa gttttccgta tgttgcatca ccttcaccct
     4741 ctccactgac agaaaatttg tgcccattaa catcaccatc taattcaaca agaattggga
     4801 caactccagt gaaaagttct tctcctttac tcatgcggta cctttctcct ctttaatgaa
     4861 ttcggtcagt gcgtcctgct gatgtgctca gtatctctat cactgatagg gatgtcaatc
     4921 tctatcactg atagggactc gaggtgaaga cgaaagggcc tcgtgctcga gtacgtctaa
     4981 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5041 cttcacctcg agaattgtga gcggataaca attgacattg tgagcggata acaagatact
     5101 gagcacatca gcaggacgca ctgaccgaat tcattaaaga ggagaaaggt accatgtcca
     5161 gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg
     5221 aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt
     5281 ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc
     5341 accatactca cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg
     5401 ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta
     5461 cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac
     5521 aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcat tttactttag
     5581 gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta
     5641 ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag
     5701 agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat
     5761 gtgaaagtgg gtcttaagct ttagcagctt aatctagagg catcaaataa aacgaaaggc
     5821 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     5881 taggacaaat ccgccgccct agacctagct gcaggtcgag gataaatatc taacaccgtg
     5941 cgtgttgact attttacctc tggcggtgat aatggttgca tgtactagaa ttcattaaag
     6001 aggagaaagg taccatatgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
     6061 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     6121 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     6181 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     6241 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     6301 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     6361 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     6421 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     6481 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     6541 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     6601 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     6661 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     6721 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     6781 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     6841 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
     6901 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     6961 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     7021 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7081 ggtttcccga ctggaaagcg ggcagtaagc ttaattagct gagtctagag gcatcaaata
     7141 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     7201 gctctcctga gtaggacaaa tccgccgccc tagacctagc tgcaggtcga gtccctatca
     7261 gtgatagaga ttgacatccc tatcagtgat agagatactg agcacatcag caggacgcac
     7321 tgaccgaatt cattaaagag gagaaaggta ccatgagcac aaaaaagaaa ccattaacac
     7381 aagagcagct tgaggacgca cgtcgcctta aagcaattta tgaaaaaaag aaaaatgaac
     7441 ttggcttatc ccaggaatct gtcgcagaca agatggggat ggggcagtca ggcgttggtg
     7501 ctttatttaa tggcatcaat gcattaaatg cttataacgc cgcattgctt gcaaaaattc
     7561 tcaaagttag cgttgaagaa tttagccctt caatcgccag agaaatctac gagatgtatg
     7621 aagcggttag tatgcagccg tcacttagaa gtgagtatga gtaccctgtt ttttctcatg
     7681 ttcaggcagg gatgttctca cctgagctta gaacctttac caaaggtgat gcggagagat
     7741 gggtaagcac aaccaaaaaa gccagtgatt ctgcattctg gcttgaggtt gaaggtaatt
     7801 ccatgaccgc accaacaggc tccaagccaa gctttcctga cggaatgtta attctcgttg
     7861 accctgagca ggctgttgag ccaggtgatt tctgcatagc cagacttggg ggtgatgagt
     7921 ttaccttcaa gaaactgatc agggatagcg gtcaggtgtt tttacaacca ctaaacccac
     7981 agtacccaat gatcccatgc aatgagagtt gttccgttgt ggggaaagtt atcgctagtc
     8041 agtggcctga agagacgttt ggctgaaaac tacgctttag cagcttgatc tagaggatcc
     8101 catggtacgc gtgctagagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     8161 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgccct
     8221 agacctaggg tacgggtttt gctgcccgca aacgggctgt tctggtgttg ctagtttgtt
     8281 atcagaatcg cagatccggc ttcaggtttg ccggctgaaa gcgctatttc ttccagaatt
     8341 gccatgattt tttccccacg ggaggcgtca ctggctcccg tgttgtcggc agctttgatt
     8401 cgataagcag catcgcctgt ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt
     8461 gtctcaggtg ttcaatttca tgttctagtt gctttgtttt actggtttca cctgttctat
     8521 taggtgttac atgctgttca tctgttacat tgtcgatctg ttcatggtga acagctttaa
     8581 atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg ttttcatctg
     8641 tgcatatgga cagttttccc tttgatatct aacggtgaac agttgttcta cttttgtttg
     8701 ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat ccttccgtat
     8761 ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga gaacgaacca
     8821 ttgagatcat gcttactttg catgtcactc aaaaattttg cctcaaaact ggtgagctga
     8881 atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttacgtag gtaggaatct
     8941 gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt ctcaagttcg
     9001 gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc agtcgggcgg
     9061 cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt acttattggt
     9121 ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag cattaacatg
     9181 aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt ttgtgttagt
     9241 tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga cttaacatgt
     9301 tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat ctcttcacta
     9361 aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg gaaaatctca
     9421 aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc tctggttgct
     9481 ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta
     9541 taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt
     9601 gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct
     9661 agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt
     9721 taatcactat accaattgag atggg
//
