LOCUS       62056_13950        5717 bp DNA     circular SYN 15-FEB-2020
DEFINITION  Expression vector pLeo1207, complete sequence.
ACCESSION   MN811118
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5717)
  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 5717)
  AUTHORS   Scheller L, Krawczyk K.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-DEC-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 
            4058, Switzerland
REFERENCE   3  (bases 1 to 5717)
  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..5717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(579..1436)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1437..1541)
                     /label=AmpR promoter
     polyA_signal    complement(1973..2197)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_RNA        complement(2365..2383)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     promoter        complement(2430..2670)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     CDS             3387..4085
                     /codon_start=1
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             4152..4748
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4916..4971
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     rep_origin      complement(join(5534..5717,1..405))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
       61 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      121 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      181 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      241 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      301 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      361 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      421 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      481 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      541 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
      601 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      661 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      721 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      781 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
      841 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
      901 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
      961 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1021 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1081 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1141 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1201 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1261 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1321 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1381 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1441 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1501 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1561 ccacctgatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa
     1621 attgtaagcg ttaatatttt gttaaaattg gatccctata cagttgaagt cggaagttta
     1681 catacactta agttggagtc attaaaactc gtttttcaac tactccacaa atttcttgtt
     1741 aacaaacaat agttttggca agtcagttag gacatctact ttgtgcatga cacaagtcat
     1801 ttttccaaca attgtttaca gacagattat ttcacttata attcactgta tcacaattcc
     1861 agtgggtcag aagtttacat acactaagtt cgactcctct gcagaatgcg gcgatgtttc
     1921 ggtaaggggt ccgctatcta gtaggcccag ctggttcttt ccgcctcaga agccatagag
     1981 cccaccgcat ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc
     2041 cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca
     2101 ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag
     2161 gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgagctcta
     2221 gacagatgcg taaggagaaa ataccgcatc agaattcaaa aaaagcaccg actcggtgcc
     2281 acttggccct gcagacatgg gtgatcctca tgttggccaa gttgataacg gactagcctt
     2341 attttaactt gctaggccct gcagacatgg gtgatcctca tgttggccta gctctaaaac
     2401 cagcccccag ccatctgccg cggtgtttcg tcctttccac aagatatata aagccaagaa
     2461 atcgaaatac tttcaagtta cggtaagcat atgatagtcc attttaaaac ataattttaa
     2521 aactgcaaac tacccaagaa attattactt tctacgtcac gtattttgta ctaatatctt
     2581 tgtgtttaca gtcaaattaa ttccaattat ctctctaaca gccttgtatc gtatatgcaa
     2641 atatgaagga atcatgggaa ataggccctc gaccggttca attgccgacc cctcccccca
     2701 acttctcggg gactgtgggc gatgtgcgct ctgcccactg acgggcaccg gagcctctag
     2761 gcgagaccct gtctcacaaa ataaagtaag cccggactga gtgcggaaag gcgggcctgg
     2821 cgggtctggt ctccccatgc gggccaccag aggccctgca gccttcagtc gcttgaaggg
     2881 gtaatggcgc ttccactcac aaacatggcg gacagagcgt gtgaacgaga tgaacagccc
     2941 ctcaaaaata tggccgccga ggctggacgg ccgtgcccca gcagcaccgc ctccgcgccc
     3001 cacgtgatct ctcgccgggc acagcgctga ccgcggaggt ccaaccggaa gaatgtccgg
     3061 attggacatt cggaagaggg cccgccttcc ctggggaatc tctgcgcacg cgcagaacgc
     3121 ttcgaccaat gaaaacacag gaagccgtcc gcgcaaccgc gttgcgtcac ttctgccgcc
     3181 cctgtttcaa ggtatatagc cgtagacgga acttcgcctt tctctcggcc ttagcgccat
     3241 ttttttgggt gagtgttttt tggttcctgc gttgggattc cgtgtacaat ccatagacat
     3301 ctgacctcgg cacttagcat catcacagca aactaactgt agcctttctc tctttccctg
     3361 tagaaacctc tgcacctgag gccaccatga gcgagctgat taaggagaac atgcacatga
     3421 agctgtacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag
     3481 gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc
     3541 ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc
     3601 acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga
     3661 gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg
     3721 acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg
     3781 tgatgcagaa gaaaacactc ggctgggagg ccttcaccga gacgctgtac cccgctgacg
     3841 gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg
     3901 caaacatcaa gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg
     3961 tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgag acctacgtcg
     4021 agcagcacga ggtggcagtg gccagatact gcgacctccc tagcaaactg gggcacaagc
     4081 tgaacggcag tggagctact aacttcagcc tgctgaagca ggctggtgac gtcgaggaga
     4141 atcctggccc catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4201 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4261 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4321 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     4381 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     4441 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     4501 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     4561 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     4621 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     4681 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     4741 ccggtgcctg attcgaaggc ctgtcgtgaa ttcactcctc aggtgcaggc tgcctatcag
     4801 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     4861 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     4921 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     4981 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     5041 acatatgcca tatgctggct gccatgaact agctactcgg gaccccttac cgaaacatcg
     5101 ccgcattctg cagaggagtc gagtgtatgt aaacttctga cccactggga atgtgatgaa
     5161 agaaataaaa gctgaaatga atcattctct ctactattat tctgatattt cacattctta
     5221 aaataaagtg gtgatcctaa ctgacctaag acagggaatt tttactagga ttaaatgtca
     5281 ggaattgtga aaaagtgagt ttaaatgtat ttggctaagg tgtatgtaaa cttccgactt
     5341 caactgtata gggatccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5401 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5461 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5521 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5581 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5641 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5701 tcccttcggg aagcgtg
//
