LOCUS       Exported               11492 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSLS51, complete sequence.
ACCESSION   KP659661
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11492
                     /organism="Cloning vector pSLS51"
                     /mol_type="other DNA"
                     /db_xref="taxon:1631297"
     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
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     CDS             1598..2392
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1601..2392
                     /codon_start=1
                     /gene="neo"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=neo
                     /note="APH 3 II; involved in neomycin resistance"
                     /protein_id="AJW72788.1"
                     /translation="MEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            1601..2392
                     /gene="neo"
                     /label=neo
     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..5351
                     /codon_start=1
                     /gene="HygEGFP"
                     /product="HygEGFP"
                     /label=HygEGFP
                     /protein_id="AJW72787.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPW
                     PTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEG
                     DTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGS
                     VQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMD
                     ELYK"
     gene            3609..5351
                     /gene="HygEGFP"
                     /label=HygEGFP
     CDS             3609..4631
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4632..5351
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     repeat_region   5522..5969
                     /label=MLV U3
                     /note="MLV U3"
     misc_recomb     5970..6003
                     /label=LE LoxP site
                     /note="LE LoxP site"
     protein_bind    complement(5970..6003)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     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"
     rep_origin      6428..6563
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             8297..8488
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     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"
     CDS             complement(9675..10538)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW"
     promoter        complement(10539..10643)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        11433..11451
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"