LOCUS       40924_28002        6670 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pleo127, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6670)
  AUTHORS   Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
  TITLE     Generalized extracellular molecule sensor platform for programming 
            cellular behavior
  JOURNAL   Nat. Chem. Biol. 14 (7), 723-729 (2018)
  PUBMED    29686358
REFERENCE   2  (bases 1 to 6670)
  AUTHORS   Scheller L, Strittmatter T.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 
            4058, Switzerland
REFERENCE   3  (bases 1 to 6670)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6670)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol."; date: "2018"; volume: "14"; issue: "7"; pages: "723-729"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058, 
            Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /label=lac operator
                     /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)."
     RBS             79..101
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    84..100
                     /label=T7_transl_en_RBS
                     /note="T7_transl_en_RBS"
     CDS             108..815
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             822..1529
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     misc_feature    822..1523
                     /label=mCherry
                     /note="mCherry"
     CDS             1551..1568
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      1635..1682
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1719..2174
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2270..3082)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      3204..3792
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3978..4120)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4225..4413)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(5188..5209)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5225..6304)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(6305..6382)
                     /label=lacI promoter
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttcccctcta gaaataattt tgtttaactt taagaaggag atatagcatg gtgagcaagg
      121 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
      181 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
      241 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
      301 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
      361 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
      421 aggacggcgg cgtggtgacc gtgacccagg actcctccct ccaggacggc gagttcatct
      481 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
      541 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
      601 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
      661 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
      721 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
      781 gccactccac cggcggcatg gacgagctgt acaagggatc cgtgagcaag ggcgaggagg
      841 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
      901 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
      961 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     1021 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     1081 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     1141 gcgtggtgac cgtgacccag gactcctccc tccaggacgg cgagttcatc tacaaggtga
     1201 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     1261 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     1321 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     1381 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     1441 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     1501 ccggcggcat ggacgagctg tacaagtaat agtaagcggc cgcactcgag caccaccacc
     1561 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1621 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1681 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     1741 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     1801 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1861 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1921 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1981 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2041 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2101 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaactagt
     2161 aacgtttaca atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2221 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     2281 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     2341 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     2401 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     2461 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     2521 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     2581 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     2641 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     2701 ggaacactgc cagcgcatca acaatgtttt cacctgaatc aggatattct tctaatacct
     2761 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     2821 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     2881 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     2941 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     3001 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     3061 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     3121 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3181 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3241 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3301 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3361 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3421 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3481 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3541 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3601 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3661 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3721 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3781 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3841 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3901 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3961 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4021 caccgcatat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     4081 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     4141 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4201 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     4261 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     4321 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     4381 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     4441 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     4501 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     4561 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     4621 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     4681 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     4741 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     4801 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     4861 cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa
     4921 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     4981 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     5041 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     5101 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     5161 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     5221 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5281 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     5341 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     5401 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     5461 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     5521 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     5581 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     5641 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     5701 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     5761 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     5821 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     5881 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     5941 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     6001 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     6061 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     6121 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     6181 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     6241 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     6301 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     6361 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     6421 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     6481 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     6541 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     6601 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     6661 tagaggatcg
//
