LOCUS       62056_14025        5818 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1261, complete sequence.
ACCESSION   MT267314
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5818)
  AUTHORS   Scheller L, Schmollack M, Bertschi A, Mansouri M, Saxena P, 
            Fussenegger M.
  TITLE     Phosphoregulated orthogonal signal transduction in mammalian cells
  JOURNAL   Nat Commun 11 (1), 3085 (2020)
  PUBMED    32555187
REFERENCE   2  (bases 1 to 5818)
  AUTHORS   Scheller L.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2020) STI IBI-STI LPDI, EPFL, AI 3123 (Batiment 
            AI), Station 19, Lausanne 1015, Switzerland
REFERENCE   3  (bases 1 to 5818)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "3085"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-MAR-2020) STI IBI-STI LPDI, EPFL, AI 3123 (Batiment AI), Station
            19, Lausanne 1015, Switzerland"
FEATURES             Location/Qualifiers
     source          1..5818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..80)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(88..104)
                     /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)."
     promoter        complement(112..142)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(157..178)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(466..1051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1225..2082)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2083..2187)
                     /label=AmpR promoter
     misc_feature    2238
                     /label=Promoter Region
                     /note="Promoter Region"
     misc_feature    2256..2459
                     /label=SV40 early promoter
                     /note="SV40 early promoter"
     promoter        2509..2527
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2559..3599
                     /codon_start=1
                     /transl_table=11
                     /product="C-ORK"
                     /label=C-ORK
                     /protein_id="QKE44353.1"
                     /translation="MTSYADALRERSHEFMNKLHVILGLLHLKSYKQLEDYILKTANNY
                     QEEIGSLLGKIKSPVIAGFLISKINRATDLGHTLILNSESQLPDSGSEDQVATLITTLG
                     NLIENALEALGPEPGGEISVTLHYRHGWLHCEVNDDGPGIAPDKIDHIFDKGVSTKGSE
                     RGVGLALVKQQVENLGGSIAVESEPGIFTQFFVQIPWDGERSNRASASGGGGSASGSQV
                     QLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQAPGKEREFLATVGWSSGITYYM
                     DSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTATRAYSVGYDYWGQGTQVTVSSA
                     SGSTGV"
     misc_feature    2568..2585
                     /label=alpha helix
                     /note="alpha helix"
     misc_feature    2586..3176
                     /label=Kinase domain 346-538
                     /note="Kinase domain 346-538"
     misc_feature    3177..3179
                     /label=DucS cytoplasmatic kinase domain
                     /note="DucS cytoplasmatic kinase domain"
     misc_feature    3207..3575
                     /label=aCaffVHH
                     /note="aCaffVHH"
     misc_feature    3294..3317
                     /label=CDR1
                     /note="CDR1"
     misc_feature    3366..3389
                     /label=CDR2
                     /note="CDR2"
     misc_feature    3507..3542
                     /label=CDR3
                     /note="CDR3"
     polyA_signal    3636..3860
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3906..4334
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4348..4677
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4744..5535
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5712..5818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
       61 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
      121 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      181 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      241 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      901 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     1021 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1081 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1141 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1201 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1261 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1321 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1381 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1441 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1501 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1561 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1621 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1681 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1741 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1801 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1861 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1921 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1981 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2041 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2101 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2161 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     2221 cggatcggga gatctccacg cgtggtaccc tcgaggcgat ctgcatctca attagtcagc
     2281 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2341 ttctccgccc catcgctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     2401 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaag
     2461 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
     2521 ctatagggag acccaagctg ttcgaagcgg aattcaccat gactagttat gctgacgcac
     2581 ttcgtgaacg atcccacgaa tttatgaata aattgcatgt gattctcgga ttattgcatc
     2641 tgaagagtta taagcagttg gaagattaca ttctcaaaac agccaataac tatcaggaag
     2701 agattggctc tctgctgggt aagatcaaat ctccggttat cgctggtttt ttaatcagca
     2761 agattaaccg cgcgaccgat ttaggccata cgctgatttt aaacagtgaa agccagctgc
     2821 cagacagcgg cagtgaggac caggtcgcga cgctgattac cacgttggga aatctgatag
     2881 aaaacgcgct ggaggcatta gggccggaac ccggaggcga aattagcgta acattgcact
     2941 accgtcacgg ctggctgcac tgtgaagtta atgatgatgg accggggatc gcacccgata
     3001 aaatcgatca catttttgac aaaggtgtct cgacaaaagg aagcgagcga ggcgtcggtt
     3061 tagcacttgt caaacaacag gtagaaaatc tcggcggcag catcgccgtg gaatcggaac
     3121 ccgggatttt cacacaattt tttgtccaga taccctggga cggggagagg tcgaacagag
     3181 ctagtgctag tggcggcggc ggcagtgcta gtggctctca ggttcaattg gtggaatctg
     3241 gagggggtct cgtacaggca ggcggttctc tccgactgag ttgcacagcc tccggtagga
     3301 ctgggaccat ctactcaatg gcctggtttc gccaggcccc aggcaaagaa agagagtttc
     3361 ttgccactgt aggttggagt tctgggatca catactacat ggattcagtt aaaggaagat
     3421 tcactatcag ccgagataaa gggaaaaata ctgtgtacct ccagatggac tctctgaaac
     3481 cggaggacac ggctgtctac tactgtacag ccacccgcgc ctactccgta gggtatgact
     3541 actgggggca aggaacacag gtaaccgtct ctagcgctag cggatccacc ggtgtctaga
     3601 aagctttgag gccggcctgc agggatcagc ctcgactgtg ccttctagtt gccagccatc
     3661 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3721 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3781 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3841 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3901 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3961 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4021 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4081 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4141 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4201 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4261 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4321 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4381 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4501 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4561 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4621 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4681 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4741 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4801 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4861 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4921 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4981 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5041 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5101 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5161 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5221 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5281 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5341 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5401 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5461 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5521 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5581 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5641 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5701 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5761 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaact
//
