LOCUS       62056_14015        5848 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1259, complete sequence.
ACCESSION   MT267312
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5848)
  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 5848)
  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 5848)
  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..5848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..861)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(862..966)
                     /label=AmpR promoter
     misc_feature    1017
                     /label=Promoter Region
                     /note="Promoter Region"
     misc_feature    1035..1238
                     /label=SV40 early promoter
                     /note="SV40 early promoter"
     promoter        1288..1306
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1338..2408
                     /codon_start=1
                     /transl_table=11
                     /product="C-ORK"
                     /label=C-ORK
                     /protein_id="QKE44351.1"
                     /translation="MTSLMQRLDGLVNYADALRERSHEFMNKLHVILGLLHLKSYKQLE
                     DYILKTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLGHTLILNSESQLPDSGSED
                     QVATLITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHCEVNDDGPGIAPDKIDHIF
                     DKGVSTKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQFFVQIPWDGERSNRASASG
                     GGGSASGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQAPGKEREFLATV
                     GWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTATRAYSVGYDYWG
                     QGTQVTVSSASGSTGV"
     misc_feature    1347..1349
                     /label=leu330
                     /note="leu330"
     misc_feature    1359..1361
                     /label=leucin 334
                     /note="leucin 334"
     misc_feature    1371..1373
                     /label=V338
                     /note="V338"
     misc_feature    1374..1376
                     /label=asn339
                     /note="asn339"
     misc_feature    1377..1394
                     /label=alpha helix
                     /note="alpha helix"
     misc_feature    1395..1988
                     /label=Kinase domain 346-538
                     /note="Kinase domain 346-538"
     misc_feature    2016..2384
                     /label=aCaffVHH
                     /note="aCaffVHH"
     misc_feature    2103..2126
                     /label=CDR1
                     /note="CDR1"
     misc_feature    2175..2198
                     /label=CDR2
                     /note="CDR2"
     misc_feature    2316..2351
                     /label=CDR3
                     /note="CDR3"
     polyA_signal    2445..2669
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2715..3143
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3157..3486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3553..4344
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4521..4654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4691..4707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4715..4731)
                     /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(4739..4769)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4784..4805)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5093..5678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctccacgc
     1021 gtggtaccct cgaggcgatc tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     1081 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atcgctgact
     1141 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     1201 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaga gctctctggc taactagaga
     1261 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgt
     1321 tcgaagcgga attcaccatg actagtctga tgcagcgact cgacggtctg gtcaactatg
     1381 ctgacgcact tcgtgaacga tcccacgaat ttatgaataa attgcatgtg attctcggat
     1441 tattgcatct gaagagttat aagcagttgg aagattacat tctcaaaaca gccaataact
     1501 atcaggaaga gattggctct ctgctgggta agatcaaatc tccggttatc gctggttttt
     1561 taatcagcaa gattaaccgc gcgaccgatt taggccatac gctgatttta aacagtgaaa
     1621 gccagctgcc agacagcggc agtgaggacc aggtcgcgac gctgattacc acgttgggaa
     1681 atctgataga aaacgcgctg gaggcattag ggccggaacc cggaggcgaa attagcgtaa
     1741 cattgcacta ccgtcacggc tggctgcact gtgaagttaa tgatgatgga ccggggatcg
     1801 cacccgataa aatcgatcac atttttgaca aaggtgtctc gacaaaagga agcgagcgag
     1861 gcgtcggttt agcacttgtc aaacaacagg tagaaaatct cggcggcagc atcgccgtgg
     1921 aatcggaacc cgggattttc acacaatttt ttgtccagat accctgggac ggggagaggt
     1981 cgaacagagc tagtgctagt ggcggcggcg gcagtgctag tggctctcag gttcaattgg
     2041 tggaatctgg agggggtctc gtacaggcag gcggttctct ccgactgagt tgcacagcct
     2101 ccggtaggac tgggaccatc tactcaatgg cctggtttcg ccaggcccca ggcaaagaaa
     2161 gagagtttct tgccactgta ggttggagtt ctgggatcac atactacatg gattcagtta
     2221 aaggaagatt cactatcagc cgagataaag ggaaaaatac tgtgtacctc cagatggact
     2281 ctctgaaacc ggaggacacg gctgtctact actgtacagc cacccgcgcc tactccgtag
     2341 ggtatgacta ctgggggcaa ggaacacagg taaccgtctc tagcgctagc ggatccaccg
     2401 gtgtctagaa agctttgagg ccggcctgca gggatcagcc tcgactgtgc cttctagttg
     2461 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2521 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2581 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2641 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2701 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2761 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2821 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2881 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2941 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3001 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3061 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3121 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3241 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3301 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3361 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3421 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3481 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3541 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3601 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3661 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3721 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3781 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3841 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3901 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3961 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4021 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4081 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4141 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4201 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4261 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4321 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4381 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4441 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4501 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4561 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4621 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4681 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4741 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4801 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4861 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4921 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4981 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5041 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5101 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5161 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5221 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5281 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5341 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5401 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5461 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5521 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5581 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     5641 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5701 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5761 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5821 gtatatatga gtaaacttgg tctgacag
//
