LOCUS Exported 7084 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral expression of shRNA targeting BNIP3. ACCESSION . VERSION . KEYWORDS pLKO-sh-BNIP3-FL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7084) TITLE Gordon Lab plasmids REFERENCE 2 (bases 1 to 7084) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7084 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 21..609 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 510..529 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 763..780 /label=L4440 /note="L4440 vector, forward primer" protein_bind 897..918 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 933..963 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 971..987 /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 976..998 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 995..1011 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 995..1011 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 1030..1050 /label=T3 /note="T3 promoter, forward primer" promoter 1032..1050 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 1078..1304 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 1305..1485 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1532..1657 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 2150..2383 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2568..2612 /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" promoter 2910..3150 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 2910..2930 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 3081..3100 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_feature 3255..3372 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 3421..3931 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 3850..3868 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 3953..4552 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3953..3972) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4449..4469 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 4680..4913 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4985..5106 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5022..5041) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5076..5095 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5146..5281 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5208..5227 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(5301..5320) /label=T7 /note="T7 promoter, forward primer" promoter complement(5302..5320) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5330..5347) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5330..5346) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5339..5361) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5488..5943 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5575..5594) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5785..5806 /label=F1ori-F /note="F1 origin, forward primer" promoter 5969..6073 /gene="bla" /label=AmpR promoter CDS 6074..6934 /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(6292..6311) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 61 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 121 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag 181 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 241 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 301 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 361 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 421 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 481 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 541 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 601 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 661 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt 721 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag 781 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa 841 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat 901 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg 961 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac 1021 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt 1081 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct 1141 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg 1201 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc 1261 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag 1321 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat 1381 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact 1441 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag 1501 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga 1561 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag 1621 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag 1681 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca 1741 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac 1801 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga 1861 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga 1921 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac 1981 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt 2041 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca 2101 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt 2161 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg 2221 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta 2281 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa 2341 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct 2401 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc 2461 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca 2521 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag 2581 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc 2641 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt 2701 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga 2761 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag 2821 agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat cgatcacgag 2881 actagcctcg agcggccgcc cccttcaccg agggcctatt tcccatgatt ccttcatatt 2941 tgcatatacg atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa 3001 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt 3061 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt 3121 tcttggcttt atatatcttg tggaaaggac gaaacaccgg aagacaccac aagataccaa 3181 cctcgaggtt ggtatcttgt ggtgtctttt tttgaattct cgacctcgag acaaatggca 3241 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga 3301 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa 3361 attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcc gcggctcgag 3421 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc 3481 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc 3541 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc 3601 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac 3661 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc 3721 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag 3781 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct 3841 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct 3901 cgttgaccga atcaccgacc tctctcccca gggggatcca ccggagctta ccatgaccga 3961 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct 4021 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat 4081 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4141 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4201 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4261 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4321 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 4381 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 4441 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 4501 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc 4561 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcgg tacctttaag 4621 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact 4681 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct 4741 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 4801 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 4861 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag 4921 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga 4981 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 5041 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 5101 atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact 5161 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 5221 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 5281 ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg 5341 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac 5401 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac 5461 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg 5521 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 5581 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 5641 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 5701 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 5761 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 5821 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 5881 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa 5941 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 6001 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 6061 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 6121 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 6181 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 6241 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 6301 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc 6361 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 6421 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 6481 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 6541 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 6601 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact 6661 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 6721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 6781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 6841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 6901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 6961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 7021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 7081 cgta //