LOCUS       62056_13955        5716 bp DNA     circular SYN 15-FEB-2020
DEFINITION  Expression vector pLeo1209, complete sequence.
ACCESSION   MN811119
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5716)
  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 5716)
  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 5716)
  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..5716
                     /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(2429..2669)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     CDS             3386..4084
                     /codon_start=1
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             4151..4747
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4915..4970
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     rep_origin      complement(join(5533..5716,1..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 tcctcgttat tgatacacac ggtgtttcgt cctttccaca agatatataa agccaagaaa
     2461 tcgaaatact ttcaagttac ggtaagcata tgatagtcca ttttaaaaca taattttaaa
     2521 actgcaaact acccaagaaa ttattacttt ctacgtcacg tattttgtac taatatcttt
     2581 gtgtttacag tcaaattaat tccaattatc tctctaacag ccttgtatcg tatatgcaaa
     2641 tatgaaggaa tcatgggaaa taggccctcg accggttcaa ttgccgaccc ctccccccaa
     2701 cttctcgggg actgtgggcg atgtgcgctc tgcccactga cgggcaccgg agcctctagg
     2761 cgagaccctg tctcacaaaa taaagtaagc ccggactgag tgcggaaagg cgggcctggc
     2821 gggtctggtc tccccatgcg ggccaccaga ggccctgcag ccttcagtcg cttgaagggg
     2881 taatggcgct tccactcaca aacatggcgg acagagcgtg tgaacgagat gaacagcccc
     2941 tcaaaaatat ggccgccgag gctggacggc cgtgccccag cagcaccgcc tccgcgcccc
     3001 acgtgatctc tcgccgggca cagcgctgac cgcggaggtc caaccggaag aatgtccgga
     3061 ttggacattc ggaagagggc ccgccttccc tggggaatct ctgcgcacgc gcagaacgct
     3121 tcgaccaatg aaaacacagg aagccgtccg cgcaaccgcg ttgcgtcact tctgccgccc
     3181 ctgtttcaag gtatatagcc gtagacggaa cttcgccttt ctctcggcct tagcgccatt
     3241 tttttgggtg agtgtttttt ggttcctgcg ttgggattcc gtgtacaatc catagacatc
     3301 tgacctcggc acttagcatc atcacagcaa actaactgta gcctttctct ctttccctgt
     3361 agaaacctct gcacctgagg ccaccatgag cgagctgatt aaggagaaca tgcacatgaa
     3421 gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg
     3481 caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc
     3541 cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca
     3601 cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca catgggagag
     3661 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga
     3721 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt
     3781 gatgcagaag aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg
     3841 cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc
     3901 aaacatcaag accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt
     3961 ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga
     4021 gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct
     4081 gaacggcagt ggagctacta acttcagcct gctgaagcag gctggtgacg tcgaggagaa
     4141 tcctggcccc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     4201 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     4261 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     4321 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     4381 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     4441 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     4501 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     4561 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     4621 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     4681 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     4741 cggtgcctga ttcgaaggcc tgtcgtgaat tcactcctca ggtgcaggct gcctatcaga
     4801 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     4861 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     4921 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     4981 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     5041 catatgccat atgctggctg ccatgaacta gctactcggg accccttacc gaaacatcgc
     5101 cgcattctgc agaggagtcg agtgtatgta aacttctgac ccactgggaa tgtgatgaaa
     5161 gaaataaaag ctgaaatgaa tcattctctc tactattatt ctgatatttc acattcttaa
     5221 aataaagtgg tgatcctaac tgacctaaga cagggaattt ttactaggat taaatgtcag
     5281 gaattgtgaa aaagtgagtt taaatgtatt tggctaaggt gtatgtaaac ttccgacttc
     5341 aactgtatag ggatccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5401 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5461 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5521 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5581 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5641 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5701 cccttcggga agcgtg
//
