LOCUS       Exported                9549 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Lentiviral vector pLenti6-eGFP-V5-Puro, complete sequence.
ACCESSION   LT009450
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9549)
  AUTHORS   De Groote P, Grootjans S, Lippens S, Eichperger C, Leurs K, Kahr I, 
            Tanghe G, Bruggeman I, De Schamphelaire W, Urwyler C, Vandenabeele 
            P, Haustraete J, Declercq W.
  TITLE     Generation of a new Gateway-compatible inducible lentiviral vector 
            platform allowing easy derivation of co-transduced cells
  JOURNAL   BioTechniques 60 (5), 252-259 (2016)
  PUBMED    27177818
REFERENCE   2  (bases 1 to 9549)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-DEC-2015) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 9549)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9549
                     /organism="Lentiviral vector pLenti6-eGFP-V5-Puro"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent University, 
                     Belgium) accession number LMBP 8402."
                     /db_xref="taxon:1783457"
     misc_feature    1..16
                     /label=HIV-1 cPPT
                     /note="HIV-1 cPPT"
     regulatory      17..251
                     /regulatory_class="terminator"
                     /note="RSV/HIV-1 dU3/3 LTR"
     LTR             18..251
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    323..444
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(323..397)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      323..397
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      484..619
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      533..608
                     /label=SV40 ori
                     /note="SV40 ori"
     promoter        complement(640..658)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(642..658)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     primer_bind     complement(668..684)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      826..1281
                     /label=f1 origin
                     /note="f1 origin"
     promoter        1307..1411
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1412..2272
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="CVH74102.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            1412..2272
                     /gene="bla"
                     /label=bla
     rep_origin      2443..3031
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2813..3215
                     /label=pMB1 ori
                     /note="pMB1 ori"
     regulatory      3307..3390
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    3319..3340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3355..3385
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3393..3409
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     3417..3433
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3454..3472
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      3454..3470
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        3500..3726
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     regulatory      3579..3907
                     /regulatory_class="promoter"
                     /note="RSV/HIV-1 5 LTR"
     LTR             3727..3907
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    3954..4079
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4018..4062
                     /label=HIV-1 psi packaging signal
                     /note="HIV-1 psi packaging signal"
     misc_feature    4572..4805
                     /label=HIV-1 Rev response element
                     /note="HIV-1 Rev response element"
     regulatory      5304..5820
                     /regulatory_class="promoter"
                     /note="hCMV-IE promoter and enhancer"
     enhancer        5313..5616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5617..5820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      complement(5937..6061)
                     /regulatory_class="attenuator"
                     /note="gateway recombination site attR1"
     protein_bind    5937..6061
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6086..6116
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             6170..6829
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyl transferase"
                     /label=cat
                     /note="chloramphenicol resistance"
                     /protein_id="CVH74103.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            6170..6829
                     /gene="cat"
                     /label=cat
     CDS             7171..7476
                     /codon_start=1
                     /gene="ccdB"
                     /product="ccdB"
                     /label=ccdB
                     /note="B gene of the control of cell death locus of the 
                     Escherichia coli F plasmid"
                     /protein_id="CVH74104.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            7171..7476
                     /gene="ccdB"
                     /label=ccdB
     regulatory      7517..7641
                     /regulatory_class="attenuator"
                     /note="gateway recombination site attR2"
     protein_bind    complement(7517..7641)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             7673..8461
                     /codon_start=1
                     /gene="EGFP"
                     /product="EGFP"
                     /label=EGFP
                     /note="Aequorea victoria green fluorescent protein,enhanced
                     red-shifted variant with V5 epitope tag"
                     /protein_id="CVH74105.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKEGPRFEGKPIPNPLLGLDSTRTG"
     gene            7673..8461
                     /gene="EGFP"
                     /label=EGFP
     CDS             7673..8389
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             8408..8449
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      8482..8764
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        8491..8821
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8672..8807
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      8721..8796
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             8873..9475
                     /codon_start=1
                     /gene="pac"
                     /product="puromycin N-acetyl-transferase"
                     /label=pac
                     /note="puromycin resistance gene"
                     /protein_id="CVH74106.1"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     gene            8873..9475
                     /gene="pac"
                     /label=pac