LOCUS       40924_44394        4255 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTSL, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4255)
  AUTHORS   Taylor NM, Prokhorov NS, Guerrero-Ferreira RC, Shneider MM, Browning
            C, Goldie KN, Stahlberg H, Leiman PG.
  TITLE     Structure of the T4 baseplate and its function in triggering sheath 
            contraction
  JOURNAL   Nature 533 (7603), 346-352 (2016)
  PUBMED    27193680
REFERENCE   2  (bases 1 to 4255)
  AUTHORS   Shneider MM, Miroshikov KA.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-DEC-2015) Laboratory of Molecular Bioengineering, 
            IBCH, Miklukho-Maklaya str. 16/10, Moscow 117997, Russian Federation
REFERENCE   3  (bases 1 to 4255)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4255)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature"; 
            date: "2016"; volume: "533"; issue: "7603"; pages: "346-352"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-DEC-2015) Laboratory of Molecular Bioengineering, IBCH, 
            Miklukho-Maklaya str. 16/10, Moscow 117997, Russian Federation"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             complement(207..227)
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             complement(786..803)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     RBS             complement(828..850)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(865..889)
                     /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(890..908)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1417..1605
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    1710..1852
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2038..2626)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2800..3657)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3658..3762)
                     /label=AmpR promoter
     rep_origin      complement(3789..4244)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgctgccc tggaaataca gattctcgcc caattcgccg
      241 ccactaccac cgctaccgcc actgccaccg ctgccaccca attgaccgtc gtggtcgtga
      301 tcgtggtggt gatcgtgcgc gccgtgaacg tgaccatgag ccagttcttc ttcagtcgct
      361 tcgcgaatcg ccacaacttc aacgttgaat ttcaggttct gaccggccag catgtggtta
      421 ccatcaacca cgacgtgatc gtcttcaacc gcagtgattt caaccggtac cggaccctgg
      481 tcggtttcag ccaggaaacg catacctacc tgcagttcat caacgcccat aaatacgtct
      541 ttaggaacac gttgcaccag gttttcgtcg tactgaccgt aagcgtcgtt cgcgccaaca
      601 gcgacatcaa atttgtcgcc aacttcatga ccttccagcg ccgtttccag gccagagatc
      661 agggaaccgt gaccatgcag gtagtccagc ggcgcactca ccggagactc atcaaccaac
      721 acaccgtctt ctgtacgtac ctgataggcc aggctgacca ccaggtcttt gcttactttg
      781 ctagcgtgat gatgatgatg atggctgctg cccatggcca tggtatatct ccttcttaaa
      841 gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag
      901 tcgtattaat ttcgcgggat cgagatctcg ggcagcgttg ggtcctggcc acgggtgcgc
      961 atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag
     1021 cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg
     1081 acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg
     1141 aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc
     1201 tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc
     1261 tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca
     1321 tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc
     1381 cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc
     1441 cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag
     1501 ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt
     1561 taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     1621 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1681 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     1741 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     1801 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     1861 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1921 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1981 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2041 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2101 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2161 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2221 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2281 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2341 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2401 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2461 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2521 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2581 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2641 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2701 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2761 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2821 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2881 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2941 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3001 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3061 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat
     3121 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3181 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3241 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3301 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3361 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3421 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3481 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3541 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3601 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3661 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3721 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3781 gccacctgaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3841 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3901 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3961 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     4021 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     4081 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4141 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     4201 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgcca
//
