LOCUS       62056_17405        6240 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pMSx5, complete sequence.
ACCESSION   MT267322
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6240)
  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 6240)
  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 6240)
  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..6240
                     /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        2696..2734
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        2779..2797
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2838..4334
                     /codon_start=1
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
                     /translation="LLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQPAQ
                     TAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDKHV
                     PDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVVTT
                     TRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRKYM
                     FRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTHLM
                     GLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESRAY
                     RALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKARD
                     RKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFARG
                     PQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     polyA_signal    4449..4673
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4719..5147
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5161..5490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             join(5557..6240,1..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 tacccctata ggggtatagc gccggctacc cctatagggg tagctgcagg tcgagggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
     2761 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg ttcgaagcgg
     2821 aattcaccat gactagtctg ctgctgctgc tgctgggcct gagactgcag ctgagcctgg
     2881 gcatcatccc cgtggaggag gagaaccccg acttctggaa cagagaggcc gccgaggccc
     2941 tgggcgccgc caagaagctg cagcccgccc agaccgccgc caagaacctg atcatcttcc
     3001 tgggcgacgg catgggcgtg agcaccgtga ccgccgccag aatcctgaag ggccagaaga
     3061 aggacaagct gggccccgag atccccctgg ccatggacag attcccctac gtggccctga
     3121 gcaagaccta caacgtggac aagcacgtgc ccgacagcgg cgccaccgcc accgcctacc
     3181 tgtgcggcgt gaagggcaac ttccagacca tcggcctgag cgccgccgcc agattcaacc
     3241 agtgcaacac caccagaggc aacgaggtga tcagcgtgat gaacagagcc aagaaggccg
     3301 gcaagagcgt gggcgtggtg accaccacca gagtgcagca cgccagcccc gccggcacct
     3361 acgcccacac cgtgaacaga aactggtaca gcgacgccga cgtgcccgcc agcgccagac
     3421 aggagggctg ccaggacatc gccacccagc tgatcagcaa catggacatc gacgtgatcc
     3481 tgggcggcgg cagaaagtac atgttcagaa tgggcacccc cgaccccgag taccccgacg
     3541 actacagcca gggcggcacc agactggacg gcaagaacct ggtgcaggag tggctggcca
     3601 agagacaggg cgccagatac gtgtggaaca gaaccgagct gatgcaggcc agcctggacc
     3661 ccagcgtgac ccacctgatg ggcctgttcg agcccggcga catgaagtac gagatccaca
     3721 gagacagcac cctggacccc agcctgatgg agatgaccga ggccgccctg agactgctga
     3781 gcagaaaccc cagaggcttc ttcctgttcg tggagggcgg cagaatcgac cacggccacc
     3841 acgagagcag agcctacaga gccctgaccg agaccatcat gttcgacgac gccatcgaga
     3901 gagccggcca gctgaccagc gaggaggaca ccctgagcct ggtgaccgcc gaccacagcc
     3961 acgtgttcag cttcggcggc taccccctga gaggcagcag catcttcggc ctggcccccg
     4021 gcaaggccag agacagaaag gcctacaccg tgctgctgta cggcaacggc cccggctacg
     4081 tgctgaagga cggcgccaga cccgacgtga ccgagagcga gagcggcagc cccgagtaca
     4141 gacagcagag cgccgtgccc ctggacgagg agacccacgc cggcgaggac gtggccgtgt
     4201 tcgccagagg cccccaggcc cacctggtgc acggcgtgca ggagcagacc ttcatcgccc
     4261 acgtgatggc cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gccccccccg
     4321 ccggcaccac cgacgccgcc caccccggct acagcagagt gggcgccgcc ggcagattcg
     4381 agcagaccgc tagcggatcc accggtgtct agaaagcttt gaggccggcc tgcagggatc
     4441 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4501 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4561 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4621 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4681 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4741 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4801 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4861 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4921 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4981 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5041 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     5101 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     5161 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5221 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5281 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5341 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5401 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5461 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5521 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5581 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5641 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5701 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5761 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5821 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5881 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5941 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6001 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6061 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     6121 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6181 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
//
