LOCUS LT009447 9870 bp DNA circular SYN 15-AUG-2016 DEFINITION Lentiviral vector pDG2i-V5-Puro, complete sequence. ACCESSION LT009447 VERSION LT009447.1 KEYWORDS . SOURCE Lentiviral vector pDG2i-V5-Puro ORGANISM Lentiviral vector pDG2i-V5-Puro other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 9870) AUTHORS De Schamphelaire W. JOURNAL Submitted (22-DEC-2015) to the INSDC. BCCM/LMBP, Universiteit Gent, Technologiepark 927, 9052, BELGIUM. REFERENCE 2 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 FEATURES Location/Qualifiers source 1..9870 /organism="Lentiviral vector pDG2i-V5-Puro" /mol_type="other DNA" /note="BCCM/LMBP Plasmid collection (Ghent University, Belgium) accession number LMBP 7946." /db_xref="taxon:1783454" misc_feature 75..90 /note="HIV-1 cPPT" regulatory 91..325 /note="RSV/HIV-1 dU3/3' LTR" /regulatory_class="terminator" regulatory 397..471 /note="SV40 polyA early" /regulatory_class="terminator" regulatory complement(397..471) /note="SV40 polyA late" /regulatory_class="terminator" rep_origin 607..682 /note="SV40 ori" regulatory complement(716..732) /note="T7 promoter" /regulatory_class="promoter" rep_origin 900..1355 /note="f1 origin" CDS 1486..2346 /transl_table=11 /gene="bla" /product="beta-lactamase" /note="ampicillin resistance" /protein_id="CVH74087.1" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" rep_origin 2887..3289 /note="pMB1 ori" regulatory 3381..3464 /note="lac promoter" /regulatory_class="promoter" regulatory 3528..3544 /note="T3 promoter" /regulatory_class="promoter" regulatory 3653..3981 /note="RSV/HIV-1 5' LTR" /regulatory_class="promoter" misc_feature 4092..4136 /note="HIV-1 psi packaging signal" misc_feature 4646..4879 /note="HIV-1 Rev response element" regulatory 5397..5664 /note="hCMVTRE promoter" /regulatory_class="promoter" regulatory complement(5707..5831) /note="gateway recombination site attR1" /regulatory_class="attenuator" CDS 5940..6599 /transl_table=11 /gene="cat" /product="chloramphenicol acetyl transferase" /note="chloramphenicol resistance" /protein_id="CVH74088.1" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG A" CDS 6941..7246 /transl_table=11 /gene="ccdB" /product="ccdB" /note="B gene of the control of cell death locus of the Escherichia coli F plasmid" /protein_id="CVH74089.1" /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSD KVSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" regulatory 7287..7411 /note="gateway recombination site attR2" /regulatory_class="attenuator" misc_feature 7464..7505 /note="V5 epitope" regulatory 7538..7820 /note="SV40 early promoter" /regulatory_class="promoter" rep_origin 7777..7852 /note="SV40 ori" CDS 7929..>8627 /transl_table=11 /gene="rtTA3" /product="Reverse tetracycline-responsive transcriptional activator" /protein_id="CVH74090.1" /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWH VKNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLG TRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERET PTTDSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLD MLPADALDDFDLDML" misc_feature 8637..8670 /note="loxP" regulatory 8693..9265 /note="5' UTR of the EMCV polyprotein gene (IRES)" /regulatory_class="ribosome_binding_site" CDS 9268..9870 /transl_table=11 /gene="pac" /product="puromycin N-acetyl-transferase" /note="puromycin resistance gene" /protein_id="CVH74091.1" /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHI ERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGS RLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAF LETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" BASE COUNT 2565 a 2392 c 2515 g 2398 t ORIGIN 1 gctaagcaca attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat 61 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga 121 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag 181 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 241 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 301 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta 361 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat 421 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 481 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc 541 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 601 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat 661 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag 721 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc 781 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag 841 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga 901 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 961 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 1021 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 1081 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 1141 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 1201 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 1261 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 1321 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg 1381 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 1441 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 1501 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 1561 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 1621 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 1681 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 1741 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 1801 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 1861 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 1921 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 1981 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 2041 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 2101 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 2161 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 2221 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 2281 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 2341 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 2401 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 2461 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 2521 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 2581 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 2641 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 2701 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 2761 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 2821 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 2881 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 2941 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 3001 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 3061 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 3121 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 3181 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 3241 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc 3301 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc 3361 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca 3421 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa 3481 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac 3541 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt 3601 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg 3661 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg 3721 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt 3781 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct 3841 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 3901 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 3961 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc 4021 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg 4081 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg 4141 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt 4201 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct 4261 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact 4321 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac 4381 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt 4441 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga 4501 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata 4561 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc 4621 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag 4681 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg 4741 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc 4801 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc 4861 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg 4921 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga 4981 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg 5041 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg 5101 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa 5161 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata 5221 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac 5281 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc 5341 gattagtgaa cggatctcga cggtatcgat ggtccgaggt tctagacgag tttactccct 5401 atcagtgata gagaacgatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga 5461 gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag 5521 aacgtatgtc gagtttatcc ctatcagtga tagagaacgt atgtcgagtt tactccctat 5581 cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag 5641 ctcgtttagt gaaccgtcag atcgcaccgg tcactagtcc agtgtggtgg aattctgcag 5701 atatcaacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat 5761 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc 5821 cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct tccggctcgt 5881 ataatgtgtg gattttgagt taggatccgg cgagattttc aggagctaag gaagctaaaa 5941 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac 6001 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata 6061 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc 6121 acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg 6181 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa 6241 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt 6301 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga 6361 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg 6421 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg 6481 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg 6541 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa 6601 gatctggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt 6661 gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtgtgctat 6721 gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca aggcatatat 6781 gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt 6841 gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa 6901 atgaacggct cttttgctga cgagaacagg gactggtgaa atgcagttta aggtttacac 6961 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac 7021 gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc 7081 ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga 7141 tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga 7201 aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccgt 7261 tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt 7321 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg 7381 tttctcgttc agctttcttg tacaaagtgg ttgatatcca gcacagtggc ggccgctcga 7441 gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt 7501 ctacgcgtac cggttagtaa tgagtttgga attaattctg tggaatgtgt gtcagttagg 7561 gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt 7621 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 7681 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 7741 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 7801 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 7861 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc 7921 acgtgaccat gtctaggctg gacaagagca aagtcataaa cggagctctg gaattactca 7981 atggtgtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc 8041 agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg 8101 agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca tggcaagact 8161 ttctgcggaa caacgccaag tcataccgct gtgctctcct ctcacatcgc gacggggcta 8221 aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa aatcagctcg 8281 cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg tccgccgtgg 8341 gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca aaagaggaaa 8401 gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt gagctgttcg 8461 accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata tgtggcctgg 8521 agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat tttgacttag 8581 acatgctccc agccgatgcc cttgacgatt ttgaccttga catgctcccc gggtaaataa 8641 cttcgtatag tatacattat acgaagttat ggatccgcgg ccgcaaattc cgcccctctc 8701 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg 8761 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg 8821 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag 8881 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt 8941 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc 9001 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga 9061 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga 9121 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct 9181 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg 9241 ttttcctttg aaaaacacga taataccatg gccaccgagt acaagcccac ggtgcgcctc 9301 gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac 9361 cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa 9421 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc 9481 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag 9541 atcggctcgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa 9601 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc 9661 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg 9721 gccgagcgcg ctggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc 9781 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc 9841 tggtgcatga cccgcaagcc cggtgcctga //