LOCUS       62056_14030        6256 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1286, complete sequence.
ACCESSION   MT267315
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6256)
  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 6256)
  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 6256)
  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..6256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..9
                     /label=SV40 polyA
                     /note="SV40 polyA"
     primer_bind     complement(49..65)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(73..89)
                     /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(97..127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(142..163)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(451..1036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1210..2067)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2068..2172)
                     /label=AmpR promoter
     misc_feature    2241..2444
                     /label=SV40 early promoter
                     /note="SV40 early promoter"
     promoter        2494..2512
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2541..2546
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             2544..4022
                     /codon_start=1
                     /transl_table=11
                     /product="Dual-ORK"
                     /label=Dual-ORK
                     /protein_id="QKE44354.1"
                     /translation="MTSGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQA
                     PGKEREFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTAT
                     RAYSVGYDYWGQGTQVTVSSASGGGGSASKTEVRKLMQRLDGLVNYADALRERSHEFMN
                     KLHVILGLLHLKSYKQLEDYILKTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLG
                     HTLILNSESQLPDSGSEDQVATLITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHC
                     EVNDDGPGIAPDKIDHIFDKGVSTKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQF
                     FVQIPWDGERSNRASASGGGGSASGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYS
                     MAWFRQAPGKEREFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTA
                     VYYCTATRAYSVGYDYWGQGTQVTVSSASGSTGV"
     misc_feature    2553..2915
                     /label=aCaffVHH
                     /note="aCaffVHH"
     misc_feature    2634..2657
                     /label=CDR1
                     /note="CDR1"
     misc_feature    2706..2729
                     /label=CDR2
                     /note="CDR2"
     misc_feature    2847..2882
                     /label=CDR3
                     /note="CDR3"
     misc_feature    2922..2936
                     /label=scfv
                     /note="scfv"
     misc_feature    2943..3602
                     /label=DcuS truncation
                     /note="DcuS truncation"
     misc_feature    3630..3998
                     /label=aCaffVHH
                     /note="aCaffVHH"
     misc_feature    3717..3740
                     /label=CDR1
                     /note="CDR1"
     misc_feature    3789..3812
                     /label=CDR2
                     /note="CDR2"
     misc_feature    3930..3965
                     /label=CDR3
                     /note="CDR3"
     polyA_signal    4059..4283
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4329..4757
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4771..5100
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5167..5958
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    6135..6256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
       61 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
      121 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
      181 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
      241 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
      301 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      361 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      421 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      481 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      541 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      601 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      661 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      721 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      781 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      841 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
      901 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      961 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     1021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1201 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1261 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1321 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1381 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1441 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1501 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1561 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1621 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1681 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1741 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1801 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1861 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1921 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1981 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2041 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2101 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2161 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     2221 ccacgcgtgg taccctcgag gcgatctgca tctcaattag tcagcaacca tagtcccgcc
     2281 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatcg
     2341 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2401 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaagagctc tctggctaac
     2461 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
     2521 agctgttcga agcggaattc accatgacta gtggctctca ggttcaattg gtggaatctg
     2581 gagggggtct cgtacaggca ggcggttctc tccgactgag ttgcacagcc tccggtagga
     2641 ctgggaccat ctactcaatg gcctggtttc gccaggcccc aggcaaagaa agagagtttc
     2701 ttgccactgt aggttggagt tctgggatca catactacat ggattcagtt aaaggaagat
     2761 tcactatcag ccgagataaa gggaaaaata ctgtgtacct ccagatggac tctctgaaac
     2821 cggaggacac ggctgtctac tactgtacag ccacccgcgc ctactccgta gggtatgact
     2881 actgggggca aggaacacag gtaaccgtct ctagcgctag tggcggcggc ggcagtgcta
     2941 gtaaaactga agtacgtaaa ctgatgcagc gactcgacgg tctggtcaac tatgctgacg
     3001 cacttcgtga acgatcccac gaatttatga ataaattgca tgtgattctc ggattattgc
     3061 atctgaagag ttataagcag ttggaagatt acattctcaa aacagccaat aactatcagg
     3121 aagagattgg ctctctgctg ggtaagatca aatctccggt tatcgctggt tttttaatca
     3181 gcaagattaa ccgcgcgacc gatttaggcc atacgctgat tttaaacagt gaaagccagc
     3241 tgccagacag cggcagtgag gaccaggtcg cgacgctgat taccacgttg ggaaatctga
     3301 tagaaaacgc gctggaggca ttagggccgg aacccggagg cgaaattagc gtaacattgc
     3361 actaccgtca cggctggctg cactgtgaag ttaatgatga tggaccgggg atcgcacccg
     3421 ataaaatcga tcacattttt gacaaaggtg tctcgacaaa aggaagcgag cgaggcgtcg
     3481 gtttagcact tgtcaaacaa caggtagaaa atctcggcgg cagcatcgcc gtggaatcgg
     3541 aacccgggat tttcacacaa ttttttgtcc agataccctg ggacggggag aggtcgaaca
     3601 gagctagtgc tagtggcggc ggcggcagtg ctagtggctc tcaggttcaa ttggtggaat
     3661 ctggaggggg tctcgtacag gcaggcggtt ctctccgact gagttgcaca gcctccggta
     3721 ggactgggac catctactca atggcctggt ttcgccaggc cccaggcaaa gaaagagagt
     3781 ttcttgccac tgtaggttgg agttctggga tcacatacta catggattca gttaaaggaa
     3841 gattcactat cagccgagat aaagggaaaa atactgtgta cctccagatg gactctctga
     3901 aaccggagga cacggctgtc tactactgta cagccacccg cgcctactcc gtagggtatg
     3961 actactgggg gcaaggaaca caggtaaccg tctctagcgc tagcggatcc accggtgtct
     4021 agaaagcttt gaggccggcc tgcagggatc agcctcgact gtgccttcta gttgccagcc
     4081 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4141 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4201 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4261 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     4321 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4381 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4441 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4501 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4561 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4621 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4681 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4741 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4801 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4861 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4921 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4981 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     5041 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5101 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5161 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5221 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5281 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5341 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5401 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5461 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5521 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5581 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5641 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5701 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5761 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5821 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5881 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5941 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6001 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6061 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6121 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6181 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6241 tcatcaatgt atctta
//
