LOCUS       62056_13965        6430 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1227, complete sequence.
ACCESSION   MT267302
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6430)
  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 6430)
  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 6430)
  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..6430
                     /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..1133
                     /label=DcuR BE2
                     /note="DcuR BE2"
     misc_feature    1152..1177
                     /label=DcuR BE1
                     /note="DcuR BE1"
     misc_feature    1191..1216
                     /label=DcuR BE2
                     /note="DcuR BE2"
     misc_feature    1235..1260
                     /label=DcuR BE1
                     /note="DcuR BE1"
     promoter        1274..1312
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        1357..1375
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1404..1409
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             1416..2912
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
     polyA_signal    3027..3251
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3297..3725
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3739..4068
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4135..4926
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5103..5236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5273..5289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5297..5313)
                     /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(5321..5351)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5366..5387)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5675..6260)
                     /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 gtgataaact ttaaaaagtg ctgcatagcg
     1141 ccggccgcgc ctgattacaa aactttaaaa agtgctggct gcaggtcgag tgattacaaa
     1201 actttaaaaa gtgctgcata gcgccggccg cgcctgatta caaaacttta aaaagtgctg
     1261 gctgcaggtc gagggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
     1321 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
     1381 ccaagctgtt cgaagcggaa ttcaccatga ctagtctgct gctgctgctg ctgggcctga
     1441 gactgcagct gagcctgggc atcatccccg tggaggagga gaaccccgac ttctggaaca
     1501 gagaggccgc cgaggccctg ggcgccgcca agaagctgca gcccgcccag accgccgcca
     1561 agaacctgat catcttcctg ggcgacggca tgggcgtgag caccgtgacc gccgccagaa
     1621 tcctgaaggg ccagaagaag gacaagctgg gccccgagat ccccctggcc atggacagat
     1681 tcccctacgt ggccctgagc aagacctaca acgtggacaa gcacgtgccc gacagcggcg
     1741 ccaccgccac cgcctacctg tgcggcgtga agggcaactt ccagaccatc ggcctgagcg
     1801 ccgccgccag attcaaccag tgcaacacca ccagaggcaa cgaggtgatc agcgtgatga
     1861 acagagccaa gaaggccggc aagagcgtgg gcgtggtgac caccaccaga gtgcagcacg
     1921 ccagccccgc cggcacctac gcccacaccg tgaacagaaa ctggtacagc gacgccgacg
     1981 tgcccgccag cgccagacag gagggctgcc aggacatcgc cacccagctg atcagcaaca
     2041 tggacatcga cgtgatcctg ggcggcggca gaaagtacat gttcagaatg ggcacccccg
     2101 accccgagta ccccgacgac tacagccagg gcggcaccag actggacggc aagaacctgg
     2161 tgcaggagtg gctggccaag agacagggcg ccagatacgt gtggaacaga accgagctga
     2221 tgcaggccag cctggacccc agcgtgaccc acctgatggg cctgttcgag cccggcgaca
     2281 tgaagtacga gatccacaga gacagcaccc tggaccccag cctgatggag atgaccgagg
     2341 ccgccctgag actgctgagc agaaacccca gaggcttctt cctgttcgtg gagggcggca
     2401 gaatcgacca cggccaccac gagagcagag cctacagagc cctgaccgag accatcatgt
     2461 tcgacgacgc catcgagaga gccggccagc tgaccagcga ggaggacacc ctgagcctgg
     2521 tgaccgccga ccacagccac gtgttcagct tcggcggcta ccccctgaga ggcagcagca
     2581 tcttcggcct ggcccccggc aaggccagag acagaaaggc ctacaccgtg ctgctgtacg
     2641 gcaacggccc cggctacgtg ctgaaggacg gcgccagacc cgacgtgacc gagagcgaga
     2701 gcggcagccc cgagtacaga cagcagagcg ccgtgcccct ggacgaggag acccacgccg
     2761 gcgaggacgt ggccgtgttc gccagaggcc cccaggccca cctggtgcac ggcgtgcagg
     2821 agcagacctt catcgcccac gtgatggcct tcgccgcctg cctggagccc tacaccgcct
     2881 gcgacctggc cccccccgcc ggcaccaccg acgccgccca ccccggctac agcagagtgg
     2941 gcgccgccgg cagattcgag cagaccgcta gcggatccac cggtgtctag aaagctttga
     3001 ggccggcctg cagggatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3061 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3121 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3181 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3241 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3301 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3361 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3421 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3481 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3541 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3601 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3661 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3721 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3841 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3901 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3961 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4021 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4081 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4141 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4201 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4261 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4321 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4381 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4441 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4501 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4561 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4621 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4681 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4741 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4801 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4861 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4921 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4981 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5041 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5101 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5161 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5221 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5281 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5341 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5401 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5461 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5521 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5581 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5641 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5701 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5761 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5821 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5881 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5941 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6001 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6061 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6121 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6181 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6241 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6301 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6361 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6421 ggtctgacag
//
