LOCUS       Exported                8369 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Lentiviral vector pDG1i-VSV-Blast, complete sequence.
ACCESSION   LT009442
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8369)
  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 8369)
  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 8369)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8369
                     /organism="Lentiviral vector pDG1i-VSV-Blast"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent University, 
                     Belgium) accession number LMBP 9592."
                     /db_xref="taxon:1783449"
     regulatory      complement(35..159)
                     /regulatory_class="attenuator"
                     /note="gateway recombination site attR1"
     protein_bind    35..159
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        184..214
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             268..927
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyl transferase"
                     /label=cat
                     /note="chloramphenicol resistance"
                     /protein_id="CVH74057.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            268..927
                     /gene="cat"
                     /label=cat
     CDS             1269..1574
                     /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="CVH74059.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            1269..1574
                     /gene="ccdB"
                     /label=ccdB
     regulatory      1615..1739
                     /regulatory_class="attenuator"
                     /note="gateway recombination site attR2"
     protein_bind    complement(1615..1739)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    1783..1815
                     /label=VSV epitope
                     /note="VSV epitope"
     CDS             1783..1815
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     regulatory      1851..2133
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        1860..2190
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2041..2176
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2090..2165
                     /label=SV40 ori
                     /note="SV40 ori"
     regulatory      2238..2303
                     /regulatory_class="promoter"
                     /note="synthetic EM7 promoter"
     promoter        2238..2285
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2304..2702
                     /codon_start=1
                     /gene="bsr"
                     /product="blasticidin S-resistance"
                     /label=bsr
                     /protein_id="CVH74061.1"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     gene            2304..2702
                     /gene="bsr"
                     /label=bsr
     misc_feature    2772..2787
                     /label=HIV-1 cPPT
                     /note="HIV-1 cPPT"
     regulatory      2788..3022
                     /regulatory_class="terminator"
                     /note="RSV/HIV-1 dU3/3 LTR"
     LTR             2789..3022
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3094..3215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(3094..3168)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      3094..3168
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      3255..3390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      3304..3379
                     /label=SV40 ori
                     /note="SV40 ori"
     promoter        complement(3411..3429)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(3413..3429)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     primer_bind     complement(3439..3455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3597..4052
                     /label=f1 origin
                     /note="f1 origin"
     promoter        4078..4182
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4183..5043
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="CVH74062.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4183..5043
                     /gene="bla"
                     /label=bla
     rep_origin      5214..5802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5584..5986
                     /label=pMB1 ori
                     /note="pMB1 ori"
     regulatory      6078..6161
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    6090..6111
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6126..6156
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6164..6180
                     /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     6188..6204
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6225..6243
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      6225..6241
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        6271..6497
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     regulatory      6350..6678
                     /regulatory_class="promoter"
                     /note="RSV/HIV-1 5 LTR"
     LTR             6498..6678
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    6725..6850
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6789..6833
                     /label=HIV-1 psi packaging signal
                     /note="HIV-1 psi packaging signal"
     misc_feature    7343..7576
                     /label=HIV-1 Rev response element
                     /note="HIV-1 Rev response element"
     protein_bind    8093..8111
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      8094..8361
                     /regulatory_class="promoter"
                     /note="hCMVTRE promoter"
     protein_bind    8128..8146
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8164..8182
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8200..8218
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8235..8253
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8271..8289
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        8301..8339
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"