LOCUS       Exported                8363 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Lentiviral vector pDG1i-HA-Blast, complete sequence.
ACCESSION   LT009439
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8363)
  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 8363)
  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 8363)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8363
                     /organism="Lentiviral vector pDG1i-HA-Blast"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent University, 
                     Belgium) accession number LMBP 9593."
                     /db_xref="taxon:1783446"
     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="CVH74036.1"
                     /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="CVH74037.1"
                     /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..1809
                     /label=HA epitope
                     /note="HA epitope"
     CDS             1783..1809
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      1845..2127
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        1854..2184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2035..2170
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2084..2159
                     /label=SV40 ori
                     /note="SV40 ori"
     regulatory      2232..2297
                     /regulatory_class="promoter"
                     /note="synthetic EM7 promoter"
     promoter        2232..2279
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2298..2696
                     /codon_start=1
                     /gene="bsr"
                     /product="blasticidin S-resistance"
                     /label=bsr
                     /protein_id="CVH74038.1"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     gene            2298..2696
                     /gene="bsr"
                     /label=bsr
     misc_feature    2766..2781
                     /label=HIV-1 cPPT
                     /note="HIV-1 cPPT"
     regulatory      2782..3016
                     /regulatory_class="terminator"
                     /note="RSV/HIV-1 dU3/3 LTR"
     LTR             2783..3016
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3088..3209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(3088..3162)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      3088..3162
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      3249..3384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      3298..3373
                     /label=SV40 ori
                     /note="SV40 ori"
     promoter        complement(3405..3423)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(3407..3423)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     primer_bind     complement(3433..3449)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3591..4046
                     /label=f1 origin
                     /note="f1 origin"
     promoter        4072..4176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4177..5037
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="CVH74039.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4177..5037
                     /gene="bla"
                     /label=bla
     rep_origin      5208..5796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5578..5980
                     /label=pMB1 ori
                     /note="pMB1 ori"
     regulatory      6072..6155
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    6084..6105
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6120..6150
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6158..6174
                     /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     6182..6198
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6219..6237
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      6219..6235
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        6265..6491
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     regulatory      6344..6672
                     /regulatory_class="promoter"
                     /note="RSV/HIV-1 5 LTR"
     LTR             6492..6672
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    6719..6844
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6783..6827
                     /label=HIV-1 psi packaging signal
                     /note="HIV-1 psi packaging signal"
     misc_feature    7337..7570
                     /label=HIV-1 Rev response element
                     /note="HIV-1 Rev response element"
     protein_bind    8087..8105
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      8088..8355
                     /regulatory_class="promoter"
                     /note="hCMVTRE promoter"
     protein_bind    8122..8140
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8158..8176
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8194..8212
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8229..8247
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8265..8283
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        8295..8333
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"