LOCUS       62056_13970        6513 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1228, complete sequence.
ACCESSION   MT267303
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6513)
  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 6513)
  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 6513)
  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..6513
                     /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    1029..1054
                     /label=DcuR BE2
                     /note="DcuR BE2"
     misc_feature    1073..1098
                     /label=DcuR BE1
                     /note="DcuR BE1"
     misc_feature    1112..1137
                     /label=DcuR BE2
                     /note="DcuR BE2"
     misc_feature    1156..1181
                     /label=DcuR BE1
                     /note="DcuR BE1"
     misc_feature    1195..1216
                     /label=DcuR BE2
                     /note="DcuR BE2"
     misc_feature    1235..1260
                     /label=DcuR BE1
                     /note="DcuR BE1"
     misc_feature    1274..1299
                     /label=DcuR BE2
                     /note="DcuR BE2"
     misc_feature    1318..1343
                     /label=DcuR BE1
                     /note="DcuR BE1"
     promoter        1357..1395
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        1440..1458
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1487..1492
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             1499..2995
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
     polyA_signal    3110..3334
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3380..3808
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3822..4151
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4218..5009
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5186..5319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5356..5372)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5380..5396)
                     /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(5404..5434)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5449..5470)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5758..6343)
                     /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 gtctcgagtg attacaaaac tttaaaaagt gctgcatagc gccggccgcg cctgattaca
     1081 aaactttaaa aagtgctggc tgcaggtcga gtgattacaa aactttaaaa agtgctgcat
     1141 agcgccggcc gcgcctgatt acaaaacttt aaaaagtgct ggctgcaggt cgagtgataa
     1201 actttaaaaa gtgctgcata gcgccggccg cgcctgatta caaaacttta aaaagtgctg
     1261 gctgcaggtc gagtgattac aaaactttaa aaagtgctgc atagcgccgg ccgcgcctga
     1321 ttacaaaact ttaaaaagtg ctggctgcag gtcgagggta ggcgtgtacg gtgggaggtc
     1381 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
     1441 aatacgactc actataggga gacccaagct gttcgaagcg gaattcacca tgactagtct
     1501 gctgctgctg ctgctgggcc tgagactgca gctgagcctg ggcatcatcc ccgtggagga
     1561 ggagaacccc gacttctgga acagagaggc cgccgaggcc ctgggcgccg ccaagaagct
     1621 gcagcccgcc cagaccgccg ccaagaacct gatcatcttc ctgggcgacg gcatgggcgt
     1681 gagcaccgtg accgccgcca gaatcctgaa gggccagaag aaggacaagc tgggccccga
     1741 gatccccctg gccatggaca gattccccta cgtggccctg agcaagacct acaacgtgga
     1801 caagcacgtg cccgacagcg gcgccaccgc caccgcctac ctgtgcggcg tgaagggcaa
     1861 cttccagacc atcggcctga gcgccgccgc cagattcaac cagtgcaaca ccaccagagg
     1921 caacgaggtg atcagcgtga tgaacagagc caagaaggcc ggcaagagcg tgggcgtggt
     1981 gaccaccacc agagtgcagc acgccagccc cgccggcacc tacgcccaca ccgtgaacag
     2041 aaactggtac agcgacgccg acgtgcccgc cagcgccaga caggagggct gccaggacat
     2101 cgccacccag ctgatcagca acatggacat cgacgtgatc ctgggcggcg gcagaaagta
     2161 catgttcaga atgggcaccc ccgaccccga gtaccccgac gactacagcc agggcggcac
     2221 cagactggac ggcaagaacc tggtgcagga gtggctggcc aagagacagg gcgccagata
     2281 cgtgtggaac agaaccgagc tgatgcaggc cagcctggac cccagcgtga cccacctgat
     2341 gggcctgttc gagcccggcg acatgaagta cgagatccac agagacagca ccctggaccc
     2401 cagcctgatg gagatgaccg aggccgccct gagactgctg agcagaaacc ccagaggctt
     2461 cttcctgttc gtggagggcg gcagaatcga ccacggccac cacgagagca gagcctacag
     2521 agccctgacc gagaccatca tgttcgacga cgccatcgag agagccggcc agctgaccag
     2581 cgaggaggac accctgagcc tggtgaccgc cgaccacagc cacgtgttca gcttcggcgg
     2641 ctaccccctg agaggcagca gcatcttcgg cctggccccc ggcaaggcca gagacagaaa
     2701 ggcctacacc gtgctgctgt acggcaacgg ccccggctac gtgctgaagg acggcgccag
     2761 acccgacgtg accgagagcg agagcggcag ccccgagtac agacagcaga gcgccgtgcc
     2821 cctggacgag gagacccacg ccggcgagga cgtggccgtg ttcgccagag gcccccaggc
     2881 ccacctggtg cacggcgtgc aggagcagac cttcatcgcc cacgtgatgg ccttcgccgc
     2941 ctgcctggag ccctacaccg cctgcgacct ggcccccccc gccggcacca ccgacgccgc
     3001 ccaccccggc tacagcagag tgggcgccgc cggcagattc gagcagaccg ctagcggatc
     3061 caccggtgtc tagaaagctt tgaggccggc ctgcagggat cagcctcgac tgtgccttct
     3121 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3181 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3241 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3301 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3361 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3421 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3481 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3541 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3601 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3661 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3721 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3781 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3841 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3901 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3961 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4021 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4081 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4141 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4201 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4261 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4321 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4381 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4441 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4501 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4561 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4621 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4681 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4741 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4801 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4861 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4921 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4981 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5041 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5101 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5161 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5221 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5281 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5341 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5401 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5461 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5521 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5581 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5641 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5701 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5761 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5821 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5881 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5941 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6001 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6061 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6121 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6181 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6241 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6301 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6361 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6421 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6481 ctaaagtata tatgagtaaa cttggtctga cag
//
