LOCUS Exported 9377 bp ds-DNA circular SYN 29-JAN-2014
DEFINITION Lentiviral Gateway(R) destination vector with a blasticidin
resistance marker and a V5 epitope tag, for strong expression and
improved plasmid yield and viral titer.
ACCESSION .
VERSION .
KEYWORDS pLX304
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9377)
AUTHORS Yang X, Boehm JS, Yang X, Salehi-Ashtiani K, Hao T, Shen Y, Lubonja
R, Thomas SR, Alkan O, Bhimdi T, Green TM, Johannessen CM, Silver
SJ, Nguyen C, Murray RR, Hieronymus H, Balcha D, Fan C, Lin C,
Ghamsari L, Vidal M, Hahn WC, Hill DE, Root DE.
TITLE A public genome-scale lentiviral expression library of human ORFs.
JOURNAL Nat. Methods 2011;8:659-61.
PUBMED 21706014
REFERENCE 2 (bases 1 to 9377)
AUTHORS Root Lab
TITLE Direct Submission
JOURNAL Exported Monday, Mar 31, 2014 from SnapGene 2.3.2
FEATURES Location/Qualifiers
source 1..9377
/organism="synthetic DNA construct"
/lab_host="Mammalian Cells"
/mol_type="other DNA"
enhancer 65..368
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 369..572
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 595..719
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 744..774
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
CDS 828..1487
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1829..2134
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/note="ccdB"
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(2175..2299)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 2301..2342
/codon_start=1
/product="epitope tag from simian virus 5"
/note="V5 tag"
/translation="GKPIPNPLLGLDST"
misc_feature 2343..2351
/note="stop codons"
misc_feature 2384..2972
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 3044..3277
/note="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 3355..3476
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 3516..3651
/note="SV40 ori"
/note="SV40 origin of replication"
rep_origin 3858..4313
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4339..4443
/gene="bla"
/note="AmpR promoter"
CDS 4444..5304
/codon_start=1
/note="AmpR"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 5475..6063
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 6530..6758
/note="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 6759..6939
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6986..7111
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7604..7837
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
promoter 8241..8741
/note="hPGK promoter"
/note="human phosphoglycerate kinase 1 (PGK1) promoter"
CDS 8753..9148
/codon_start=1
/gene="Aspergillusterreus BSD"
/product="blasticidin S deaminase"
/note="BSD"
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
misc_feature 9208..9325
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 cccggggttattaatagtaatcaattacggggtcattagttcatagcccatatatggagt
61 tccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcc
121 cattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgac
181 gtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcata
241 tgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgccc
301 agtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgcta
361 ttaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcac
421 ggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatc
481 aacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggc
541 gtgtacggtgggaggtctatataagcagagctctctggctaactgtcgggatcaacaagt
601 ttgtacaaaaaagctgaacgagaaacgtaaaatgatataaatatcaatatattaaattag
661 attttgcataaaaaacagactacataatactgtaaaacacaacatatccagtcactatgg
721 cggccgcattaggcaccccaggctttacactttatgcttccggctcgtataatgtgtgga
781 ttttgagttaggatccgtcgagattttcaggagctaaggaagctaaaatggagaaaaaaa
841 tcactggatataccaccgttgatatatcccaatggcatcgtaaagaacattttgaggcat
901 ttcagtcagttgctcaatgtacctataaccagaccgttcagctggatattacggcctttt
961 taaagaccgtaaagaaaaataagcacaagttttatccggcctttattcacattcttgccc
1021 gcctgatgaatgctcatccggaattccgtatggcaatgaaagacggtgagctggtgatat
1081 gggatagtgttcacccttgttacaccgttttccatgagcaaactgaaacgttttcatcgc
1141 tctggagtgaataccacgacgatttccggcagtttctacacatatattcgcaagatgtgg
1201 cgtgttacggtgaaaacctggcctatttccctaaagggtttattgagaatatgtttttcg
1261 tctcagccaatccctgggtgagtttcaccagttttgatttaaacgtggccaatatggaca
1321 acttcttcgcccccgttttcaccatgggcaaatattatacgcaaggcgacaaggtgctga
1381 tgccgctggcgattcaggttcatcatgccgtttgtgatggcttccatgtcggcagaatgc
1441 ttaatgaattacaacagtactgcgatgagtggcagggcggggcgtaaagatctggatccg
1501 gcttactaaaagccagataacagtatgcgtatttgcgcgctgatttttgcggtataagaa
1561 tatatactgatatgtatacccgaagtatgtcaaaaagaggtatgctatgaagcagcgtat
1621 tacagtgacagttgacagcgacagctatcagttgctcaaggcatatatgatgtcaatatc
1681 tccggtctggtaagcacaaccatgcagaatgaagcccgtcgtctgcgtgccgaacgctgg
1741 aaagcggaaaatcaggaagggatggctgaggtcgcccggtttattgaaatgaacggctct
1801 tttgctgacgagaacaggggctggtgaaatgcagtttaaggtttacacctataaaagaga
1861 gagccgttatcgtctgtttgtggatgtacagagtgatattattgacacgcccgggcgacg
1921 gatggtgatccccctggccagtgcacgtctgctgtcagataaagtctcccgtgaacttta
1981 cccggtggtgcatatcggggatgaaagctggcgcatgatgaccaccgatatggccagtgt
2041 gccggtctccgttatcggggaagaagtggctgatctcagccaccgcgaaaatgacatcaa
2101 aaacgccattaacctgatgttctggggaatataaatgtcaggctcccttatacacagcca
2161 gtctgcaggtcgaccatagtgactggatatgttgtgttttacagtattatgtagtctgtt
2221 ttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcag
2281 ctttcttgtacaaagtggttggtaagcctatccctaaccctctcctcggtctcgattcta
2341 cgtagtaatgagctagcgctaaccggtggcgcgttaagtcgacaatcaacctctggatta
2401 caaaatttgtgaaagattgactggtattcttaactatgttgctccttttacgctatgtgg
2461 atacgctgctttaatgcctttgtatcatgctattgcttcccgtatggctttcattttctc
2521 ctccttgtataaatcctggttgctgtctctttatgaggagttgtggcccgttgtcaggca
2581 acgtggcgtggtgtgcactgtgtttgctgacgcaacccccactggttggggcattgccac
2641 cacctgtcagctcctttccgggactttcgctttccccctccctattgccacggcggaact
2701 catcgccgcctgccttgcccgctgctggacaggggctcggctgttgggcactgacaattc
2761 cgtggtgttgtcggggaaatcatcgtcctttccttggctgctcgcctgtgttgccacctg
2821 gattctgcgcgggacgtccttctgctacgtcccttcggccctcaatccagcggaccttcc
2881 ttcccgcggcctgctgccggctctgcggcctcttccgcgtcttcgccttcgccctcagac
2941 gagtcggatctccctttgggccgcctccccgcgtcgactttaagaccaatgacttacaag
3001 gcagctgtagatcttagccactttttaaaagaaaaggggggactggaagggctaattcac
3061 tcccaacgaagacaagatctgctttttgcttgtactgggtctctctggttagaccagatc
3121 tgagcctgggagctctctggctaactagggaacccactgcttaagcctcaataaagcttg
3181 ccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtgactctggtaactagagatcc
3241 ctcagacccttttagtcagtgtggaaaatctctagcagtacgtatagtagttcatgtcat
3301 cttattattcagtatttataacttgcaaagaaatgaatatcagagagtgagaggaacttg
3361 tttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaataaa
3421 gcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcat
3481 gtctggctctagctatcccgcccctaactccgcccatcccgcccctaactccgcccagtt
3541 ccgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggccg
3601 cctcggcctctgagctattccagaagtagtgaggaggcttttttggaggcctagggacgt
3661 acccaattcgccctatagtgagtcgtattacgcgcgctcactggccgtcgttttacaacg
3721 tcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatcccccttt
3781 cgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcag
3841 cctgaatggcgaatgggacgcgccctgtagcggcgcattaagcgcggcgggtgtggtggt
3901 tacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttctt
3961 cccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccc
4021 tttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtga
4081 tggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtc
4141 cacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggt
4201 ctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagct
4261 gatttaacaaaaatttaacgcgaattttaacaaaatattaacgcttacaatttaggtggc
4321 acttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaat
4381 atgtatccgctcatgagacaataaccctgataaatgcttcaataatattgaaaaaggaag
4441 agtatgagtattcaacatttccgtgtcgcccttattcccttttttgcggcattttgcctt
4501 cctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggt
4561 gcacgagtgggttacatcgaactggatctcaacagcggtaagatccttgagagttttcgc
4621 cccgaagaacgttttccaatgatgagcacttttaaagttctgctatgtggcgcggtatta
4681 tcccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgac
4741 ttggttgagtactcaccagtcacagaaaagcatcttacggatggcatgacagtaagagaa
4801 ttatgcagtgctgccataaccatgagtgataacactgcggccaacttacttctgacaacg
4861 atcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgtaactcgc
4921 cttgatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacg
4981 atgcctgtagcaatggcaacaacgttgcgcaaactattaactggcgaactacttactcta
5041 gcttcccggcaacaattaatagactggatggaggcggataaagttgcaggaccacttctg
5101 cgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtggg
5161 tctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatc
5221 tacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggt
5281 gcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagatt
5341 gatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctc
5401 atgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaag
5461 atcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaa
5521 aaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccg
5581 aaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtag
5641 ttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctg
5701 ttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacga
5761 tagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagc
5821 ttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgcc
5881 acgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacagga
5941 gagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggttt
6001 cgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatgg
6061 aaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcac
6121 atgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtga
6181 gctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcg
6241 gaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagc
6301 tggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagt
6361 tagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgt
6421 ggaattgtgagcggataacaatttcacacaggaaacagctatgaccatgattacgccaag
6481 cgcgcaattaaccctcactaaagggaacaaaagctggagctgcaagcttaatgtagtctt
6541 atgcaatactcttgtagtcttgcaacatggtaacgatgagttagcaacatgccttacaag
6601 gagagaaaaagcaccgtgcatgccgattggtggaagtaaggtggtacgatcgtgccttat
6661 taggaaggcaacagacgggtctgacatggattggacgaaccactgaattgccgcattgca
6721 gagatattgtatttaagtgcctagctcgatacataaacgggtctctctggttagaccaga
6781 tctgagcctgggagctctctggctaactagggaacccactgcttaagcctcaataaagct
6841 tgccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtgactctggtaactagagat
6901 ccctcagacccttttagtcagtgtggaaaatctctagcagtggcgcccgaacagggactt
6961 gaaagcgaaagggaaaccagaggagctctctcgacgcaggactcggcttgctgaagcgcg
7021 cacggcaagaggcgaggggcggcgactggtgagtacgccaaaaattttgactagcggagg
7081 ctagaaggagagagatgggtgcgagagcgtcagtattaagcgggggagaattagatcgcg
7141 atgggaaaaaattcggttaaggccagggggaaagaaaaaatataaattaaaacatatagt
7201 atgggcaagcagggagctagaacgattcgcagttaatcctggcctgttagaaacatcaga
7261 aggctgtagacaaatactgggacagctacaaccatcccttcagacaggatcagaagaact
7321 tagatcattatataatacagtagcaaccctctattgtgtgcatcaaaggatagagataaa
7381 agacaccaaggaagctttagacaagatagaggaagagcaaaacaaaagtaagaccaccgc
7441 acagcaagcggccgctgatcttcagacctggaggaggagatatgagggacaattggagaa
7501 gtgaattatataaatataaagtagtaaaaattgaaccattaggagtagcacccaccaagg
7561 caaagagaagagtggtgcagagagaaaaaagagcagtgggaataggagctttgttccttg
7621 ggttcttgggagcagcaggaagcactatgggcgcagcgtcaatgacgctgacggtacagg
7681 ccagacaattattgtctggtatagtgcagcagcagaacaatttgctgagggctattgagg
7741 cgcaacagcatctgttgcaactcacagtctggggcatcaagcagctccaggcaagaatcc
7801 tggctgtggaaagatacctaaaggatcaacagctcctggggatttggggttgctctggaa
7861 aactcatttgcaccactgctgtgccttggaatgctagttggagtaataaatctctggaac
7921 agatttggaatcacacgacctggatggagtgggacagagaaattaacaattacacaagct
7981 taatacactccttaattgaagaatcgcaaaaccagcaagaaaagaatgaacaagaattat
8041 tggaattagataaatgggcaagtttgtggaattggtttaacataacaaattggctgtggt
8101 atataaaattattcataatgatagtaggaggcttggtaggtttaagaatagtttttgctg
8161 tactttctatagtgaatagagttaggcagggatattcaccattatcgtttcagacccacc
8221 tcccaaccccgaggggacccttgcgccttttccaaggcagccctgggtttgcgcagggac
8281 gcggctgctctgggcgtggttccgggaaacgcagcggcgccgaccctgggtctcgcacat
8341 tcttcacgtccgttcgcagcgtcacccggatcttcgccgctacccttgtgggccccccgg
8401 cgacgcttcctgctccgcccctaagtcgggaaggttccttgcggttcgcggcgtgccgga
8461 cgtgacaaacggaagccgcacgtctcactagtaccctcgcagacggacagcgccagggag
8521 caatggcagcgcgccgaccgcgatgggctgtggccaatagcggctgctcagcagggcgcg
8581 ccgagagcagcggccgggaaggggcggtgcgggaggcggggtgtggggcggtagtgtggg
8641 ccctgttcctgcccgcgcggtgttccgcattctgcaagcctccggagcgcacgtcggcag
8701 tcggctccctcgttgaccgaatcaccgacctctctccccagggggtaccaccatggccaa
8761 gcctttgtctcaagaagaatccaccctcattgaaagagcaacggctacaatcaacagcat
8821 ccccatctctgaagactacagcgtcgccagcgcagctctctctagcgacggccgcatctt
8881 cactggtgtcaatgtatatcattttactgggggaccttgtgcagaactcgtggtgctggg
8941 cactgctgctgctgcggcagctggcaacctgacttgtatcgtcgcgatcggaaatgagaa
9001 caggggcatcttgagcccctgcggacggtgccgacaggtgcttctcgatctgcatcctgg
9061 gatcaaagccatagtgaaggacagtgatggacagccgacggcagttgggattcgtgaatt
9121 gctgccctctggttatgtgtgggagggcctgcagctgcagtagtaagaattctagatctt
9181 gagacaaatggcagtattcatccacaattttaaaagaaaaggggggattggggggtacag
9241 tgcaggggaaagaatagtagacataatagcaacagacatacaaactaaagaattacaaaa
9301 acaaattacaaaaattcaaaattttcgggtttattacagggacagcagagatccactttg
9361 gcgccggctcgaggggg
//