LOCUS       62056_13960        6347 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1216, complete sequence.
ACCESSION   MT267301
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6347)
  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 6347)
  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 6347)
  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..6347
                     /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..1050
                     /label=DcuR BE2
                     /note="DcuR BE2"
     misc_feature    1069..1094
                     /label=DcuR BE1
                     /note="DcuR BE1"
     misc_feature    1108..1133
                     /label=DcuR BE2
                     /note="DcuR BE2"
     misc_feature    1152..1177
                     /label=DcuR BE1
                     /note="DcuR BE1"
     promoter        1191..1229
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        1274..1292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1321..1326
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             1333..2829
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
     polyA_signal    2944..3168
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3214..3642
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3656..3985
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4052..4843
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5020..5153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5190..5206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5214..5230)
                     /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(5238..5268)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5283..5304)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5592..6177)
                     /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 ataaacttta aaaagtgctg catagcgccg gccgcgcctg attacaaaac
     1081 tttaaaaagt gctggctgca ggtcgagtga ttacaaaact ttaaaaagtg ctgcatagcg
     1141 ccggccgcgc ctgattacaa aactttaaaa agtgctggct gcaggtcgag ggtaggcgtg
     1201 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
     1261 ggcttatcga aattaatacg actcactata gggagaccca agctgttcga agcggaattc
     1321 accatgacta gtctgctgct gctgctgctg ggcctgagac tgcagctgag cctgggcatc
     1381 atccccgtgg aggaggagaa ccccgacttc tggaacagag aggccgccga ggccctgggc
     1441 gccgccaaga agctgcagcc cgcccagacc gccgccaaga acctgatcat cttcctgggc
     1501 gacggcatgg gcgtgagcac cgtgaccgcc gccagaatcc tgaagggcca gaagaaggac
     1561 aagctgggcc ccgagatccc cctggccatg gacagattcc cctacgtggc cctgagcaag
     1621 acctacaacg tggacaagca cgtgcccgac agcggcgcca ccgccaccgc ctacctgtgc
     1681 ggcgtgaagg gcaacttcca gaccatcggc ctgagcgccg ccgccagatt caaccagtgc
     1741 aacaccacca gaggcaacga ggtgatcagc gtgatgaaca gagccaagaa ggccggcaag
     1801 agcgtgggcg tggtgaccac caccagagtg cagcacgcca gccccgccgg cacctacgcc
     1861 cacaccgtga acagaaactg gtacagcgac gccgacgtgc ccgccagcgc cagacaggag
     1921 ggctgccagg acatcgccac ccagctgatc agcaacatgg acatcgacgt gatcctgggc
     1981 ggcggcagaa agtacatgtt cagaatgggc acccccgacc ccgagtaccc cgacgactac
     2041 agccagggcg gcaccagact ggacggcaag aacctggtgc aggagtggct ggccaagaga
     2101 cagggcgcca gatacgtgtg gaacagaacc gagctgatgc aggccagcct ggaccccagc
     2161 gtgacccacc tgatgggcct gttcgagccc ggcgacatga agtacgagat ccacagagac
     2221 agcaccctgg accccagcct gatggagatg accgaggccg ccctgagact gctgagcaga
     2281 aaccccagag gcttcttcct gttcgtggag ggcggcagaa tcgaccacgg ccaccacgag
     2341 agcagagcct acagagccct gaccgagacc atcatgttcg acgacgccat cgagagagcc
     2401 ggccagctga ccagcgagga ggacaccctg agcctggtga ccgccgacca cagccacgtg
     2461 ttcagcttcg gcggctaccc cctgagaggc agcagcatct tcggcctggc ccccggcaag
     2521 gccagagaca gaaaggccta caccgtgctg ctgtacggca acggccccgg ctacgtgctg
     2581 aaggacggcg ccagacccga cgtgaccgag agcgagagcg gcagccccga gtacagacag
     2641 cagagcgccg tgcccctgga cgaggagacc cacgccggcg aggacgtggc cgtgttcgcc
     2701 agaggccccc aggcccacct ggtgcacggc gtgcaggagc agaccttcat cgcccacgtg
     2761 atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcccc ccccgccggc
     2821 accaccgacg ccgcccaccc cggctacagc agagtgggcg ccgccggcag attcgagcag
     2881 accgctagcg gatccaccgg tgtctagaaa gctttgaggc cggcctgcag ggatcagcct
     2941 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3001 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3061 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3121 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3181 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3241 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3301 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3361 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3421 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3481 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3541 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3601 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3661 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3721 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3781 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3841 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3901 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3961 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4021 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4081 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4141 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4201 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4261 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4321 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4381 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4441 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4501 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4561 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4621 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4681 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4741 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4801 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4861 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4921 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4981 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5041 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5101 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5161 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5221 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5281 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5341 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5401 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5461 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5521 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5581 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5641 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5701 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5761 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5821 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5881 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5941 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6001 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6061 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6121 ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6181 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6241 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6301 agttttaaat caatctaaag tatatatgag taaacttggt ctgacag
//
