pSLS51 vector (V003144)

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LOCUS       40924_40672       11492 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSLS51, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11492)
  AUTHORS   Sanber KS, Knight SB, Stephen SL, Bailey R, Escors D, Minshull J, 
            Santilli G, Thrasher AJ, Collins MK, Takeuchi Y.
  TITLE     Construction of stable packaging cell lines for clinical lentiviral 
            vector production
  JOURNAL   Sci Rep 5, 9021 (2015)
  PUBMED    25762005
REFERENCE   2  (bases 1 to 11492)
  AUTHORS   Sanber KS, Knight SB, Stephen SL, Bailey R, Escors D, Minshull J, 
            Santilli G, Thrasher AJ, Collins MK, Takeuchi Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JAN-2015) UCL Division of Infection and Immunity, 
            University College London, Gower Street, London WC1E 6BT, UK
REFERENCE   3  (bases 1 to 11492)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11492)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep 5, 
            9021 (2015)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-JAN-2015) UCL Division of Infection and Immunity, University 
            College London, Gower Street, London WC1E 6BT, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        314..517
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    738..937
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             1138..1554
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     CDS             1598..2389
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     enhancer        2874..3253
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3254..3457
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             3609..4631
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4632..5348
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     repeat_region   5522..5969
                     /label=MLV U3
                     /note="MLV U3"
     protein_bind    5970..6003
                     /label=lox66
                     /note="Right element (RE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   6013..6080
                     /label=MLV R
                     /note="MLV R"
     repeat_region   6081..6157
                     /label=MLV U5
                     /note="MLV U5"
     promoter        complement(6414..6743)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             8297..8485
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    8590..8730
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(8916..9504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(9973..10354)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     promoter        complement(10539..10643)
                     /label=AmpR promoter
     promoter        11433..11451
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"