LOCUS       62056_13485        4533 bp DNA     circular SYN 15-FEB-2020
DEFINITION  Expression vector pKK89, complete sequence.
ACCESSION   MN811113
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4533)
  AUTHORS   Krawczyk K, Scheller L, Kim H, Fussenegger M.
  TITLE     Rewiring of endogenous signaling pathways to genomic targets for 
            therapeutic cell reprogramming
  JOURNAL   Nat Commun 11 (1), 608 (2020)
  PUBMED    32001704
REFERENCE   2  (bases 1 to 4533)
  AUTHORS   Krawczyk KK.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-DEC-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 
            4058, Switzerland
REFERENCE   3  (bases 1 to 4533)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "608"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-DEC-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058, 
            Switzerland"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(178..766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(940..1797)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1798..1902)
                     /label=AmpR promoter
     rep_origin      1929..2384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    2515..2563
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    2577..2668
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
     promoter        2846..2877
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    2916..2966
                     /gene="NanoLuc-Fc"
                     /label=Gaussia luciferase signal peptide
                     /note="Gaussia luciferase signal peptide"
     CDS             2967..3476
                     /label=Nluc
                     /note="NanoLuc(R) luciferase"
     misc_feature    3477..3497
                     /gene="NanoLuc-Fc"
                     /label=flexible linker
                     /note="flexible linker"
     misc_feature    3498..4160
                     /gene="NanoLuc-Fc"
                     /label=murine Fc
                     /note="murine Fc"
     polyA_signal    complement(4189..4310)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
       61 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
      121 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      181 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      241 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      301 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
      361 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
      421 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      481 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      541 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
      601 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
      661 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
      721 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
      781 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      841 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
      901 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
      961 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1021 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1081 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1141 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1201 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     1261 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     1321 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1381 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     1441 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     1501 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     1561 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     1621 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     1681 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     1741 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     1801 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1861 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     1921 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1981 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2041 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2101 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2161 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2221 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2281 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2341 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcccatt cgccattcag
     2401 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagcccaa
     2461 gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg ccgcaataaa
     2521 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
     2581 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
     2641 gcaagtgcag gtgccagaac atttctctat cgataggtac ctgagctcgc tagctacatt
     2701 ggaaaatttt atacacgttc tagctacatt ggaaaatttt atacacgttc tagctacatt
     2761 ggaaaatttt atacacgttc tagctacatt ggaaaatttt atacacgttc tagctacatt
     2821 ggaaaatttt atacacgtta gactctagag ggtatataat ggaagctcga cttccagctt
     2881 ggcaatccgg tactgttggt aaagaattcg ccaccatggg cgtcaaggtg ctgttcgccc
     2941 tcatttgtat agctgtcgct gaggcggtct tcacactcga agatttcgtt ggggactggc
     3001 gacagacagc cggctacaac ctggaccaag tccttgaaca gggaggtgtg tccagtttgt
     3061 ttcagaatct cggggtgtcc gtaactccga tccaaaggat tgtcctgagc ggtgaaaatg
     3121 ggctgaagat cgacatccat gtcatcatcc cgtatgaagg tctgagcggc gaccaaatgg
     3181 gccagatcga aaaaattttt aaggtggtgt accctgtgga tgatcatcac tttaaggtga
     3241 tcctgcacta tggcacactg gtaatcgacg gggttacgcc gaacatgatc gactatttcg
     3301 gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa gatcactgta acagggaccc
     3361 tgtggaacgg caacaaaatt atcgacgagc gcctgatcaa ccccgacggc tccctgctgt
     3421 tccgagtaac catcaacgga gtgaccggct ggcggctgtg cgaacgcatt ctggcgggag
     3481 gcggtggaag tggtggttgt aagccttgca tatgtacagt cccagaagta tcatctgtct
     3541 tcatcttccc cccaaagccc aaggatgtgc tcaccattac tctgactcct aaggtcacgt
     3601 gtgttgtggt agacatcagc aaggatgatc ccgaggtcca gttcagctgg tttgtagatg
     3661 atgtggaggt gcacacagct cagacgcaac cccgggagga gcagttcaac agcactttcc
     3721 gctcagtcag tgaacttccc atcatgcacc aggactggct caatggcaag gagttcaaat
     3781 gcagggtcaa cagtgcagct ttccctgccc ccatcgagaa aaccatctcc aaaaccaaag
     3841 gcagaccgaa ggctccacag gtgtacacca ttccacctcc caaggagcag atggccaagg
     3901 ataaagtcag tctgacctgc atgataacag acttcttccc tgaagacatt actgtggagt
     3961 ggcagtggaa tgggcagcca gcggagaact acaagaacac tcagcccatc atggacacag
     4021 atggctctta cttcgtctac agcaagctca atgtgcagaa gagcaactgg gaggcaggaa
     4081 atactttcac ctgctctgtg ttacatgagg gcctgcacaa ccaccatact gagaagagcc
     4141 tctcccactc tcctggtaaa tgaggccggc cgcttcgagc agacatgata agatacattg
     4201 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4261 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4321 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     4381 aaaacctcta caaatgtggg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     4441 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     4501 caactcgtag gacaggtgcc ggcagcgctc ttc
//
