LOCUS Exported 7754 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION 3rd gen lentiviral empty control vector, CMV promoter, Blast . ACCESSION . VERSION . KEYWORDS pLenti CMV Blast empty (w263-1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7754) AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD. TITLE A versatile viral system for expression and depletion of proteins in mammalian cells. JOURNAL PLoS One. 2009 Aug 6;4(8):e6529. PUBMED 19657394 REFERENCE 2 (bases 1 to 7754) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7754 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 5..22 /label=L4440 /note="L4440 vector, forward primer" protein_bind 139..160 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 175..205 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 213..229 /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 218..240 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 237..253 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 237..253 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 272..292 /label=T3 /note="T3 promoter, forward primer" promoter 274..292 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 320..546 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 547..727 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 774..899 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1392..1625 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1810..1854 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2121..2237 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 2260..2563 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2564..2767 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2718..2738 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2764..2788 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 2872..2896 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2965..2989) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_feature 3020..3608 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3073..3093) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3491..3502) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3659..3679) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3664..3993 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3844..3979 /label=SV40 ori /note="SV40 origin of replication" promoter 4041..4088 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4107..4505 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" LTR 4592..4825 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4897..5018 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4934..4953) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4988..5007 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5058..5193 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5213..5232) /label=T7 /note="T7 promoter, forward primer" promoter complement(5214..5232) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5242..5259) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5242..5258) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5251..5273) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5400..5855 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5487..5506) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5697..5718 /label=F1ori-F /note="F1 origin, forward primer" promoter 5881..5985 /gene="bla" /label=AmpR promoter CDS 5986..6846 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6204..6223) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7017..7605 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7506..7525 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc 61 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg 121 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca 181 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg 241 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa 301 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta 361 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg 421 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt 481 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac 541 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 601 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 661 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 721 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc 781 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga 841 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca 901 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa 961 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag 1021 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac 1081 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct 1141 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg 1201 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga 1261 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt 1321 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga 1381 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc 1441 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag 1501 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg 1561 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag 1621 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat 1681 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg 1741 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac 1801 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat 1861 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc 1921 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga 1981 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa 2041 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga 2101 tctcgacggt atcggttaac ttttaaaaga aaagggggga ttggggggta cagtgcaggg 2161 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt 2221 acaaaattca aaattttatc gataagcttg ggagttccgc gttacataac ttacggtaaa 2281 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt 2341 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta 2401 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt 2461 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc 2521 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca 2581 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat 2641 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa 2701 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag 2761 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct 2821 ccatagaaga caccgactct agtccagtgt ggtggaattc tgcagatatc aacaagtttg 2881 tacaaaaaag caggctccac catgggaacc aattcagtcg actggatccg gtaccgaatt 2941 cgcggccgca ctcgagatat ctagacccag ctttcttgta caaagtggtt gatatccagc 3001 acagtggcgg ccgctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg 3061 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 3121 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 3181 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 3241 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 3301 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 3361 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac 3421 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 3481 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 3541 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc 3601 ctccccgcct ggaattctgc agatatccgg ttagtaatga gtttggaatt aattctgtgg 3661 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 3721 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 3781 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 3841 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 3901 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 3961 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 4021 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat 4081 aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa gaatccaccc 4141 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg 4201 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta 4261 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca 4321 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac 4381 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg 4441 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg 4501 gctaagcaca attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat 4561 cttagccact ttttaaaaga aaagggggga ctggaagggc tagctcactc ccaacgaaga 4621 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag 4681 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 4741 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 4801 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta 4861 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat 4921 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 4981 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc 5041 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 5101 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat 5161 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag 5221 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc 5281 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag 5341 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga 5401 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 5461 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 5521 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 5581 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 5641 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 5701 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 5761 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 5821 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg 5881 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 5941 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 6001 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 6061 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 6121 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 6181 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 6241 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 6301 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 6361 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 6421 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 6481 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 6541 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 6601 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 6661 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 6721 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 6781 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 6841 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 6901 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 6961 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 7021 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 7081 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 7141 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 7201 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 7261 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 7321 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 7381 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 7441 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 7501 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 7561 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 7621 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 7681 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 7741 agccgaacga ccga //