LOCUS LT009444 9846 bp DNA circular SYN 15-AUG-2016
DEFINITION Lentiviral vector pDG2i-Flag-Puro, complete sequence.
ACCESSION LT009444
VERSION LT009444.1
KEYWORDS .
SOURCE Lentiviral vector pDG2i-Flag-Puro
ORGANISM Lentiviral vector pDG2i-Flag-Puro
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 9846)
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..9846
/organism="Lentiviral vector pDG2i-Flag-Puro"
/mol_type="other DNA"
/note="BCCM/LMBP Plasmid collection (Ghent University,
Belgium) accession number LMBP 7944."
/db_xref="taxon:1783451"
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="CVH74069.1"
/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="CVH74070.1"
/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="CVH74071.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSD
KVSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
regulatory 7287..7411
/note="gateway recombination site attR2"
/regulatory_class="attenuator"
misc_feature 7455..7478
/note="FLAG epitope"
regulatory 7514..7796
/note="SV40 early promoter"
/regulatory_class="promoter"
rep_origin 7753..7828
/note="SV40 ori"
CDS 7905..>8603
/transl_table=11
/gene="rtTA3"
/product="Reverse tetracycline-responsive transcriptional
activator"
/protein_id="CVH74073.1"
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWH
VKNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLG
TRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERET
PTTDSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLD
MLPADALDDFDLDML"
misc_feature 8613..8646
/note="loxP"
regulatory 8669..9241
/note="5' UTR of the EMCV polyprotein gene (IRES)"
/regulatory_class="ribosome_binding_site"
CDS 9244..9846
/transl_table=11
/gene="pac"
/product="puromycin N-acetyl-transferase"
/note="puromycin resistance gene"
/protein_id="CVH74074.1"
/translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHI
ERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGS
RLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAF
LETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
BASE COUNT 2568 a 2379 c 2512 g 2387 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 cccggactac aaggatgacg atgacaaaac gcgtaccggt tagtaatgag
7501 tttggaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
7561 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
7621 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
7681 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
7741 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
7801 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
7861 cgggagcttg tatatccatt ttcggatctg atcagcacgt gaccatgtct aggctggaca
7921 agagcaaagt cataaacgga gctctggaat tactcaatgg tgtcggtatc gaaggcctga
7981 cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac tggcacgtga
8041 agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg catcataccc
8101 acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac gccaagtcat
8161 accgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc acccgcccaa
8221 cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct
8281 ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg ggctgcgtat
8341 tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc accgattcta
8401 tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc gaacctgcct
8461 tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag tgcgaaagcg
8521 gcgggccgac cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg
8581 acgattttga ccttgacatg ctccccgggt aaataacttc gtatagtata cattatacga
8641 agttatggat ccgcggccgc aaattccgcc cctctccctc ccccccccct aacgttactg
8701 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
8761 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
8821 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
8881 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
8941 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
9001 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
9061 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
9121 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
9181 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataat
9241 accatggcca ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg
9301 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac
9361 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
9421 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
9481 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gctcgcgcat ggccgagttg
9541 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
9601 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
9661 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgctgg ggtgcccgcc
9721 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
9781 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
9841 gcctga
//