LOCUS       62056_21580        6450 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pTS1017, complete sequence.
ACCESSION   MT267334
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6450)
  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 6450)
  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 6450)
  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..6450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    957..1181
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1227..1655
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1669..1998
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2065..2856
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3033..3166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3203..3219)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3227..3243)
                     /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(3251..3281)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3296..3317)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3605..4190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4364..5221)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5222..5326)
                     /label=AmpR promoter
     protein_bind    complement(5624..5642)
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        5654..5692
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        5737..5755
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5796..6450,1..842)
                     /codon_start=1
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
                     /translation="LLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQPAQ
                     TAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDKHV
                     PDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVVTT
                     TRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRKYM
                     FRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTHLM
                     GLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESRAY
                     RALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKARD
                     RKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFARG
                     PQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
ORIGIN
        1 gaaagtacat gttcagaatg ggcacccccg accccgagta ccccgacgac tacagccagg
       61 gcggcaccag actggacggc aagaacctgg tgcaggagtg gctggccaag agacagggcg
      121 ccagatacgt gtggaacaga accgagctga tgcaggccag cctggacccc agcgtgaccc
      181 acctgatggg cctgttcgag cccggcgaca tgaagtacga gatccacaga gacagcaccc
      241 tggaccccag cctgatggag atgaccgagg ccgccctgag actgctgagc agaaacccca
      301 gaggcttctt cctgttcgtg gagggcggca gaatcgacca cggccaccac gagagcagag
      361 cctacagagc cctgaccgag accatcatgt tcgacgacgc catcgagaga gccggccagc
      421 tgaccagcga ggaggacacc ctgagcctgg tgaccgccga ccacagccac gtgttcagct
      481 tcggcggcta ccccctgaga ggcagcagca tcttcggcct ggcccccggc aaggccagag
      541 acagaaaggc ctacaccgtg ctgctgtacg gcaacggccc cggctacgtg ctgaaggacg
      601 gcgccagacc cgacgtgacc gagagcgaga gcggcagccc cgagtacaga cagcagagcg
      661 ccgtgcccct ggacgaggag acccacgccg gcgaggacgt ggccgtgttc gccagaggcc
      721 cccaggccca cctggtgcac ggcgtgcagg agcagacctt catcgcccac gtgatggcct
      781 tcgccgcctg cctggagccc tacaccgcct gcgacctggc cccccccgcc ggcaccaccg
      841 acgccgccca ccccggctac agcagagtgg gcgccgccgg cagattcgag cagaccgcta
      901 gcggatccac cggtgtctag aaagctttga ggccggcctg cagggatcag cctcgactgt
      961 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1021 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1081 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1141 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     1201 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1261 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1321 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1381 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1441 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1501 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1561 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1621 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     1681 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1741 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1801 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     1861 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     1921 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     1981 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     2041 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2101 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2161 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2221 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     2281 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2341 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2401 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     2461 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     2521 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2581 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     2641 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     2701 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     2761 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     2821 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     2881 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     2941 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3001 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3061 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3121 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3181 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3241 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3301 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3361 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3421 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3481 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3541 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3601 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3661 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3721 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3781 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3841 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3901 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3961 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4021 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4081 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4141 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4201 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4261 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4321 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4381 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4441 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4501 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4561 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4621 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4681 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4741 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4801 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4861 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4921 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4981 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5041 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5101 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5161 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5221 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5281 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5341 aagtgccacc tgacgtcgac ggatcgggag atctccacgc gtccgtacac gcctaaagca
     5401 tatacgttct ctatcactga tagggagtaa actggatata cgttctctat cactgatagg
     5461 gagtaaactg tagatacgtt ctctatcact gatagggagt aaactggtca tacgttctct
     5521 atcactgata gggagtaaac tccttatacg ttctctatca ctgataggga gtaaagtctg
     5581 catacgttct ctatcactga tagggagtaa actcttcata cgttctctat cactgatagg
     5641 gagtaaactc gagggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
     5701 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
     5761 ccaagctgtt cgaagcggaa ttcaccatga ctagtctgct gctgctgctg ctgggcctga
     5821 gactgcagct gagcctgggc atcatccccg tggaggagga gaaccccgac ttctggaaca
     5881 gagaggccgc cgaggccctg ggcgccgcca agaagctgca gcccgcccag accgccgcca
     5941 agaacctgat catcttcctg ggcgacggca tgggcgtgag caccgtgacc gccgccagaa
     6001 tcctgaaggg ccagaagaag gacaagctgg gccccgagat ccccctggcc atggacagat
     6061 tcccctacgt ggccctgagc aagacctaca acgtggacaa gcacgtgccc gacagcggcg
     6121 ccaccgccac cgcctacctg tgcggcgtga agggcaactt ccagaccatc ggcctgagcg
     6181 ccgccgccag attcaaccag tgcaacacca ccagaggcaa cgaggtgatc agcgtgatga
     6241 acagagccaa gaaggccggc aagagcgtgg gcgtggtgac caccaccaga gtgcagcacg
     6301 ccagccccgc cggcacctac gcccacaccg tgaacagaaa ctggtacagc gacgccgacg
     6361 tgcccgccag cgccagacag gagggctgcc aggacatcgc cacccagctg atcagcaaca
     6421 tggacatcga cgtgatcctg ggcggcggca
//
