LOCUS       Exported                8382 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Lentiviral vector pDG1i-V5-Blast, complete sequence.
ACCESSION   LT009441
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8382)
  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 8382)
  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 8382)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8382
                     /organism="Lentiviral vector pDG1i-V5-Blast"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent University, 
                     Belgium) accession number LMBP 9591."
                     /db_xref="taxon:1783448"
     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="CVH74052.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            268..927
                     /gene="cat"
                     /label=cat
     CDS             1267..1572
                     /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="CVH74054.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            1267..1572
                     /gene="ccdB"
                     /label=ccdB
     regulatory      1613..1737
                     /regulatory_class="attenuator"
                     /note="gateway recombination site attR2"
     protein_bind    complement(1613..1737)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    1790..1831
                     /label=V5 epitope
                     /note="V5 epitope"
     CDS             1790..1831
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      1864..2146
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        1873..2203
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2054..2189
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2103..2178
                     /label=SV40 ori
                     /note="SV40 ori"
     regulatory      2251..2316
                     /regulatory_class="promoter"
                     /note="synthetic EM7 promoter"
     promoter        2251..2298
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2317..2715
                     /codon_start=1
                     /gene="bsr"
                     /product="blasticidin S-resistance"
                     /label=bsr
                     /protein_id="CVH74055.1"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     gene            2317..2715
                     /gene="bsr"
                     /label=bsr
     misc_feature    2785..2800
                     /label=HIV-1 cPPT
                     /note="HIV-1 cPPT"
     regulatory      2801..3035
                     /regulatory_class="terminator"
                     /note="RSV/HIV-1 dU3/3 LTR"
     LTR             2802..3035
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3107..3228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(3107..3181)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      3107..3181
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      3268..3403
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      3317..3392
                     /label=SV40 ori
                     /note="SV40 ori"
     promoter        complement(3424..3442)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(3426..3442)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     primer_bind     complement(3452..3468)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3610..4065
                     /label=f1 origin
                     /note="f1 origin"
     promoter        4091..4195
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4196..5056
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="CVH74056.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4196..5056
                     /gene="bla"
                     /label=bla
     rep_origin      5227..5815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5597..5999
                     /label=pMB1 ori
                     /note="pMB1 ori"
     regulatory      6091..6174
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    6103..6124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6139..6169
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6177..6193
                     /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     6201..6217
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6238..6256
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      6238..6254
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        6284..6510
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     regulatory      6363..6691
                     /regulatory_class="promoter"
                     /note="RSV/HIV-1 5 LTR"
     LTR             6511..6691
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    6738..6863
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6802..6846
                     /label=HIV-1 psi packaging signal
                     /note="HIV-1 psi packaging signal"
     misc_feature    7356..7589
                     /label=HIV-1 Rev response element
                     /note="HIV-1 Rev response element"
     protein_bind    8106..8124
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      8107..8374
                     /regulatory_class="promoter"
                     /note="hCMVTRE promoter"
     protein_bind    8141..8159
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8177..8195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8213..8231
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8248..8266
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8284..8302
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        8314..8352
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"