LOCUS V043285 8953 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043285 VERSION V043285 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /codon_start=1 /note="recognized by the 2H10 single-chain llama nanobody" /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /transl_table=1 /translation="KNEQELLELDKWASL" /label="gp41 peptide" misc_feature 1804..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3807 /label="Slc7a11(NM_001107673)" /note="Slc7a11(NM_001107673)" /gene="Slc7a11" CDS 3814..3879 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3923..4134 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4147..4415 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4442..5041 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" misc_feature 5048..5636 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5710..5943 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6015..6136 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6155..6290 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6328..6344) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6352..6368 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(6376..6406) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6421..6442 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6730..7318) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7489..8349) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(8350..8454) /gene="bla" /label="AmpR promoter" primer_bind 8928..8944 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggtcaga aagccagttg tggccaccat ctccaaagga 2341 ggttacctgc agggcaatgt gagcgggagg ctcccctccg tgggggacca agagccacct 2401 gggcatgaga aggtggttct gaaaaagaag atcactttgc tgaggggggt ctccatcatc 2461 atcggcaccg tcatcggatc gggcatcttc atctccccca agggcatact ccagaacacg 2521 ggcagcgtgg gcatgtcact ggtgttctgg tctgcctgtg gagtactgtc actttttgga 2581 gccctgtctt atgctgaatt gggtacgagc ataaagaaat ctggtggtca ttacacatac 2641 attctggagg tctttggtcc cttgctagct tttgttcgag tctgggtgga actgctggta 2701 atacgccccg gagctacggc tgtgatatcc ctggcttttg gacgctacat tctagaaccg 2761 ttttttattc aatgtgaaat tcctgaactt gcaatcaagc ttgtaacagc tgtgggcatc 2821 actgtggtga tggttctaaa tagcacgagt gtcagctgga gtgcccggat ccagattttc 2881 ctaacctttt gcaagctcac agcaattctg ataattatag tccctggagt tatacagcta 2941 attaaagggc aaacacatca ctttaaagat gcattttcag gaagagatac aaatctaatg 3001 gggttgccct tggcttttta ttacgggatg tatgcatatg ctggctggtt ttacctcaac 3061 tttattactg aagaagtaga caaccctgaa aaaaccatcc cccttgcaat ctgcatctct 3121 atggccatca tcacagttgg ctatgtcctg acaaatgtgg cctattttac aaccattagc 3181 gccgaggagc tgttgcagtc cagcgctgtg gcggtgacct tctctgagcg gctgctggga 3241 aaattctcat tagcagtccc gatctttgtt gccctctcct gcttcggctc catgaacggt 3301 ggtgtgtttg ctgtctccag gttattctat gttgcatctc gagaagggca ccttccggaa 3361 atcctctcca tgattcacgt ccacaagcac actcctctgc cagctgttat tgttttgcat 3421 cctctgacaa tgataatgct cttctccgga gacctctaca gtcttctgaa tttcctcagt 3481 tttgccaggt ggctttttat gggcctggca gtcgccgggc tgatttatct tcgatacaaa 3541 cgcccagata tgcatcgtcc tttcaaggtg cctctgttca tcccagcatt attctccttc 3601 acctgcctct tcatggttgt cctctccctt tactcggatc cgtttagcac cggggttggc 3661 ttccttatca ccttgactgg ggtcccggcg tattacctct tcattgtatg ggacaagaaa 3721 cccaagtggt tcagacgatt gtcagacaga ataaccagaa cattacagat tatactagaa 3781 gttgtaccag aagactctaa agaattagga tccgactaca aagaccatga cggtgattat 3841 aaagatcatg acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc 3901 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 3961 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 4021 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 4081 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 4141 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 4201 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 4261 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 4321 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 4381 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 4441 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt 4501 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga 4561 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga 4621 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga 4681 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg 4741 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga 4801 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg 4861 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct 4921 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc 4981 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg 5041 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5101 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5161 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5221 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 5281 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 5341 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 5401 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg 5461 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 5521 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 5581 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 5641 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 5701 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta 5761 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 5821 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 5881 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 5941 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 6001 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 6061 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 6121 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc 6181 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 6241 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg 6301 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 6361 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6421 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6481 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6541 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6601 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 6661 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6721 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6781 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6841 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6901 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 6961 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7021 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7081 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7141 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7201 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7261 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7321 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7381 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7441 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7501 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7561 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7621 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7681 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7741 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7801 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7861 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7921 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 7981 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8041 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8101 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8161 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8221 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8281 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8341 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8401 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8461 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8521 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 8581 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 8641 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 8701 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 8761 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 8821 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 8881 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 8941 cagtgccaag ctg //