LOCUS       62056_21585        6292 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pTS566, complete sequence.
ACCESSION   MT267333
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6292)
  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 6292)
  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 6292)
  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..6292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    285..418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(455..471)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(479..495)
                     /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(503..533)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(548..569)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(857..1442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1616..2473)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2474..2578)
                     /label=AmpR promoter
     promoter        2748..2786
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        2831..2849
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2890..4386
                     /codon_start=1
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
                     /translation="LLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQPAQ
                     TAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDKHV
                     PDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVVTT
                     TRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRKYM
                     FRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTHLM
                     GLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESRAY
                     RALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKARD
                     RKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFARG
                     PQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     polyA_signal    4501..4725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4771..5199
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5213..5542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             join(5609..6292,1..108)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
ORIGIN
        1 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
       61 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
      121 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
      181 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
      241 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
      301 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
      361 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
      421 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
      481 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
      541 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
      601 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
      661 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      721 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      781 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      841 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1021 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1261 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1381 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1441 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1501 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1561 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1621 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1681 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1741 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1801 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1861 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1921 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1981 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2041 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2101 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2161 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2221 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2281 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2341 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2401 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2461 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2521 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2581 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctccac gcgtctcgag
     2641 atttcccgta aatcgtcgag tggacatttc ccgtaaatcg tcgagtggac atttcccgta
     2701 aatcgtcgag tggacatttc ccgtaaatcg tcgagctgca ggtcgagggt aggcgtgtac
     2761 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
     2821 ttatcgaaat taatacgact cactataggg agacccaagc tgttcgaagc ggaattcacc
     2881 atgactagtc tgctgctgct gctgctgggc ctgagactgc agctgagcct gggcatcatc
     2941 cccgtggagg aggagaaccc cgacttctgg aacagagagg ccgccgaggc cctgggcgcc
     3001 gccaagaagc tgcagcccgc ccagaccgcc gccaagaacc tgatcatctt cctgggcgac
     3061 ggcatgggcg tgagcaccgt gaccgccgcc agaatcctga agggccagaa gaaggacaag
     3121 ctgggccccg agatccccct ggccatggac agattcccct acgtggccct gagcaagacc
     3181 tacaacgtgg acaagcacgt gcccgacagc ggcgccaccg ccaccgccta cctgtgcggc
     3241 gtgaagggca acttccagac catcggcctg agcgccgccg ccagattcaa ccagtgcaac
     3301 accaccagag gcaacgaggt gatcagcgtg atgaacagag ccaagaaggc cggcaagagc
     3361 gtgggcgtgg tgaccaccac cagagtgcag cacgccagcc ccgccggcac ctacgcccac
     3421 accgtgaaca gaaactggta cagcgacgcc gacgtgcccg ccagcgccag acaggagggc
     3481 tgccaggaca tcgccaccca gctgatcagc aacatggaca tcgacgtgat cctgggcggc
     3541 ggcagaaagt acatgttcag aatgggcacc cccgaccccg agtaccccga cgactacagc
     3601 cagggcggca ccagactgga cggcaagaac ctggtgcagg agtggctggc caagagacag
     3661 ggcgccagat acgtgtggaa cagaaccgag ctgatgcagg ccagcctgga ccccagcgtg
     3721 acccacctga tgggcctgtt cgagcccggc gacatgaagt acgagatcca cagagacagc
     3781 accctggacc ccagcctgat ggagatgacc gaggccgccc tgagactgct gagcagaaac
     3841 cccagaggct tcttcctgtt cgtggagggc ggcagaatcg accacggcca ccacgagagc
     3901 agagcctaca gagccctgac cgagaccatc atgttcgacg acgccatcga gagagccggc
     3961 cagctgacca gcgaggagga caccctgagc ctggtgaccg ccgaccacag ccacgtgttc
     4021 agcttcggcg gctaccccct gagaggcagc agcatcttcg gcctggcccc cggcaaggcc
     4081 agagacagaa aggcctacac cgtgctgctg tacggcaacg gccccggcta cgtgctgaag
     4141 gacggcgcca gacccgacgt gaccgagagc gagagcggca gccccgagta cagacagcag
     4201 agcgccgtgc ccctggacga ggagacccac gccggcgagg acgtggccgt gttcgccaga
     4261 ggcccccagg cccacctggt gcacggcgtg caggagcaga ccttcatcgc ccacgtgatg
     4321 gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggccccccc cgccggcacc
     4381 accgacgccg cccaccccgg ctacagcaga gtgggcgccg ccggcagatt cgagcagacc
     4441 gctagcggat ccaccggtgt ctagaaagct ttgaggccgg cctgcaggga tcagcctcga
     4501 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4561 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4621 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4681 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     4741 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     4801 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4861 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4921 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4981 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5041 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5101 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5161 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     5221 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5281 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5341 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5401 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5461 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     5521 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     5581 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     5641 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5701 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5761 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     5821 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5881 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5941 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     6001 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     6061 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6121 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6181 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6241 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata tt
//
