LOCUS       62056_14020        5836 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1260, complete sequence.
ACCESSION   MT267313
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5836)
  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 5836)
  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 5836)
  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..5836
                     /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..3617
                     /codon_start=1
                     /transl_table=11
                     /product="C-ORK"
                     /label=C-ORK
                     /protein_id="QKE44352.1"
                     /translation="MTSLDGLVNYADALRERSHEFMNKLHVILGLLHLKSYKQLEDYIL
                     KTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLGHTLILNSESQLPDSGSEDQVAT
                     LITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHCEVNDDGPGIAPDKIDHIFDKGV
                     STKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQFFVQIPWDGERSNRASASGGGGS
                     ASGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQAPGKEREFLATVGWSS
                     GITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTATRAYSVGYDYWGQGTQ
                     VTVSSASGSTGV"
     misc_feature    2568..2570
                     /label=leucin 334
                     /note="leucin 334"
     misc_feature    2580..2582
                     /label=V338
                     /note="V338"
     misc_feature    2583..2585
                     /label=asn339
                     /note="asn339"
     misc_feature    2586..2603
                     /label=alpha helix
                     /note="alpha helix"
     misc_feature    2604..3197
                     /label=Kinase domain 346-538
                     /note="Kinase domain 346-538"
     misc_feature    3225..3593
                     /label=aCaffVHH
                     /note="aCaffVHH"
     misc_feature    3312..3335
                     /label=CDR1
                     /note="CDR1"
     misc_feature    3384..3407
                     /label=CDR2
                     /note="CDR2"
     misc_feature    3525..3560
                     /label=CDR3
                     /note="CDR3"
     polyA_signal    3654..3878
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3924..4352
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4366..4695
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4762..5553
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5730..5836
                     /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 gactagtctc gacggtctgg
     2581 tcaactatgc tgacgcactt cgtgaacgat cccacgaatt tatgaataaa ttgcatgtga
     2641 ttctcggatt attgcatctg aagagttata agcagttgga agattacatt ctcaaaacag
     2701 ccaataacta tcaggaagag attggctctc tgctgggtaa gatcaaatct ccggttatcg
     2761 ctggtttttt aatcagcaag attaaccgcg cgaccgattt aggccatacg ctgattttaa
     2821 acagtgaaag ccagctgcca gacagcggca gtgaggacca ggtcgcgacg ctgattacca
     2881 cgttgggaaa tctgatagaa aacgcgctgg aggcattagg gccggaaccc ggaggcgaaa
     2941 ttagcgtaac attgcactac cgtcacggct ggctgcactg tgaagttaat gatgatggac
     3001 cggggatcgc acccgataaa atcgatcaca tttttgacaa aggtgtctcg acaaaaggaa
     3061 gcgagcgagg cgtcggttta gcacttgtca aacaacaggt agaaaatctc ggcggcagca
     3121 tcgccgtgga atcggaaccc gggattttca cacaattttt tgtccagata ccctgggacg
     3181 gggagaggtc gaacagagct agtgctagtg gcggcggcgg cagtgctagt ggctctcagg
     3241 ttcaattggt ggaatctgga gggggtctcg tacaggcagg cggttctctc cgactgagtt
     3301 gcacagcctc cggtaggact gggaccatct actcaatggc ctggtttcgc caggccccag
     3361 gcaaagaaag agagtttctt gccactgtag gttggagttc tgggatcaca tactacatgg
     3421 attcagttaa aggaagattc actatcagcc gagataaagg gaaaaatact gtgtacctcc
     3481 agatggactc tctgaaaccg gaggacacgg ctgtctacta ctgtacagcc acccgcgcct
     3541 actccgtagg gtatgactac tgggggcaag gaacacaggt aaccgtctct agcgctagcg
     3601 gatccaccgg tgtctagaaa gctttgaggc cggcctgcag ggatcagcct cgactgtgcc
     3661 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3721 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3781 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3841 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3901 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3961 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4021 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4081 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4141 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4201 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4261 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4321 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4381 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4441 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4501 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4561 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4621 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4681 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4741 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4801 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4861 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4921 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4981 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5041 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5101 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5161 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5221 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5281 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5341 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5401 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5461 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5521 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5581 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5641 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5701 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5761 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5821 gtggtttgtc caaact
//
