LOCUS       40924_28092        3400 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo665, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3400)
  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 3400)
  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 3400)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3400)
  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..3400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    15..285
                     /label=tetracycline response element
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     misc_feature    318..442
                     /label=CMVmin
                     /note="CMVmin"
     promoter        318..355
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     sig_peptide     467..526
                     /label=Ig-kappa leader
                     /note="leader sequence from mouse immunoglobulin kappa 
                     light
                     chain"
     CDS             545..1057
                     /codon_start=1
                     /label=Nluc
                     /note="NanoLuc(R) luciferase"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    complement(1087..1221)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1583..2171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2638..3016)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     promoter        complement(3201..3305)
                     /label=AmpR promoter
ORIGIN
        1 ctcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
       61 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
      121 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
      181 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
      241 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
      301 ctcggtaccc gggtcgagta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt
      361 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      421 ccgggaccga tccagcctcc gcggccccga attcaccatg actagtgaga cagacacact
      481 cctgctatgg gtactgctgc tctgggttcc aggttccact ggtgacgcta gtggtggttc
      541 tggtatggtc ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa
      601 cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc
      661 cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca
      721 tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt
      781 taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact atggcacact
      841 ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat
      901 cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat
      961 tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg
     1021 agtgaccggc tggcggctgt gcgaacgcat tctggcggct agcggatcca ccggtgtcta
     1081 gccttcgatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1141 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1201 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1261 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     1321 tatgatcctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     1381 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     1441 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     1501 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     1561 gtcgggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1621 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1681 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1741 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1801 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1861 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1921 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1981 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     2041 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2101 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2161 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2221 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2281 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2341 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2401 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2461 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2521 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2581 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtgcgcaac
     2641 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2701 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2761 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2821 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2881 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2941 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3001 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3061 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3121 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3181 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3241 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3301 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     3361 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
