LOCUS       Exported                9098 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Lentiviral vector pDG1i-eGFP-V5-Blast, complete sequence.
ACCESSION   LT009438
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9098)
  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 9098)
  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 9098)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9098
                     /organism="Lentiviral vector pDG1i-eGFP-V5-Blast"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent University, 
                     Belgium) accession number LMBP 7941"
                     /db_xref="taxon:1783445"
     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="CVH74026.1"
                     /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="CVH74027.1"
                     /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"
     CDS             1771..2559
                     /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="CVH74029.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKEGPRFEGKPIPNPLLGLDSTRTG"
     gene            1771..2559
                     /gene="EGFP"
                     /label=EGFP
     CDS             1771..2487
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2506..2547
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      2580..2862
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        2589..2919
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2770..2905
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2819..2894
                     /label=SV40 ori
                     /note="SV40 ori"
     regulatory      2967..3032
                     /regulatory_class="promoter"
                     /note="synthetic EM7 promoter"
     promoter        2967..3014
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3033..3431
                     /codon_start=1
                     /gene="bsr"
                     /product="blasticidin S-resistance"
                     /label=bsr
                     /protein_id="CVH74031.1"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     gene            3033..3431
                     /gene="bsr"
                     /label=bsr
     misc_feature    3501..3516
                     /label=HIV-1 cPPT
                     /note="HIV-1 cPPT"
     regulatory      3517..3751
                     /regulatory_class="terminator"
                     /note="RSV/HIV-1 dU3/3 LTR"
     LTR             3518..3751
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3823..3944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(3823..3897)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      3823..3897
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     rep_origin      3984..4119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4033..4108
                     /label=SV40 ori
                     /note="SV40 ori"
     promoter        complement(4140..4158)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(4142..4158)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     primer_bind     complement(4168..4184)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4326..4781
                     /label=f1 origin
                     /note="f1 origin"
     promoter        4807..4911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4912..5772
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="CVH74034.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4912..5772
                     /gene="bla"
                     /label=bla
     rep_origin      5943..6531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6313..6715
                     /label=pMB1 ori
                     /note="pMB1 ori"
     regulatory      6807..6890
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    6819..6840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6855..6885
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6893..6909
                     /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     6917..6933
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6954..6972
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      6954..6970
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        7000..7226
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     regulatory      7079..7407
                     /regulatory_class="promoter"
                     /note="RSV/HIV-1 5 LTR"
     LTR             7227..7407
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    7454..7579
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7518..7562
                     /label=HIV-1 psi packaging signal
                     /note="HIV-1 psi packaging signal"
     misc_feature    8072..8305
                     /label=HIV-1 Rev response element
                     /note="HIV-1 Rev response element"
     protein_bind    8822..8840
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      8823..9090
                     /regulatory_class="promoter"
                     /note="hCMVTRE promoter"
     protein_bind    8857..8875
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8893..8911
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8929..8947
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8964..8982
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9000..9018
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        9030..9068
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"