LOCUS V045153 9013 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045153 VERSION V045153 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..3867 /label="PLIN1(NM_002666)" /note="PLIN1(NM_002666)" /gene="PLIN1" CDS 3874..3939 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3983..4194 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4207..4475 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4502..5101 /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 5108..5696 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5770..6003 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6075..6196 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6215..6350 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6388..6404) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6412..6428 /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(6436..6466) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6481..6502 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6790..7378) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7549..8409) /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(8410..8514) /gene="bla" /label="AmpR promoter" primer_bind 8988..9004 /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 catggcagtc aacaaaggcc tcaccttgct ggatggagac 2341 ctccctgagc aggagaatgt gctgcagcgg gtcctgcagc tgccggtggt gagtggcacc 2401 tgcgaatgct tccagaagac ctacaccagc actaaggaag cccaccccct ggtggcctct 2461 gtgtgcaatg cctatgagaa gggcgtgcag agcgccagta gcttggctgc ctggagcatg 2521 gagccggtgg tccgcaggct gtccacccag ttcacagctg ccaatgagct ggcctgccga 2581 ggcttggacc acctggagga aaagatcccc gccctccagt acccccctga aaagattgct 2641 tctgagctga aggacaccat ctccacccgc ctccgcagtg ccagaaacag catcagcgtt 2701 cccatcgcga gcacttcaga caaggtcctg ggggccgctt tggccgggtg cgagcttgcc 2761 tggggggtgg ccagagacac tgcggaattt gctgccaaca ctcgagctgg ccgactggct 2821 tctggagggg ccgacttggc cttgggcagc attgagaagg tggtggagta cctcctccct 2881 ccagacaagg aagagtcagc ccctgctcct ggacaccagc aagcccagaa gtctcccaag 2941 gccaagccaa gcctcttgag cagggttggg gctctgacca acaccctctc tcgatacacc 3001 gtgcagacca tggcccgggc cctggagcag ggccacaccg tggccatgtg gatcccaggc 3061 gtggtgcccc tgagcagcct ggcccagtgg ggtgcctcag tggccatgca ggcggtgtcc 3121 cggcggagga gcgaagtgcg ggtaccctgg ctgcacagcc tcgcagccgc ccaggaggag 3181 gatcatgagg accagacaga cacggaggga gaggacacgg aggaggagga agaattggag 3241 actgaggaga acaagttcag tgaggtagca gccctgccag gccctcgagg cctcctgggt 3301 ggtgtggcac ataccctgca gaagaccctc cagaccacca tctcggctgt gacatgggca 3361 cctgcagctg tgctgggcat ggcagggagg gtgctgcacc tcacaccagc ccctgctgtc 3421 tcctcaacca aggggagggc catgtcccta tcagatgccc tgaagggcgt tactgacaac 3481 gtggtggaca cagtggtgca ttacgtgccg ctccccaggc tgtcgctgat ggagcccgag 3541 agcgaattcc gggacatcga caacccacca gccgaggtcg agcgccggga ggcggagcgc 3601 agagcgtctg gggcgccgtc cgccggcccg gagcccgccc cgcgtctcgc acagccccgc 3661 cgcagcctgc gcagcgcgca gagccccggc gcgccccccg gcccgggcct ggaggacgaa 3721 gtcgccacgc ccgcagcgcc gcgcccgggc ttcccggccg tgccccgcga gaagccaaag 3781 cgcagggtca gcgacagctt cttccggccc agcgtcatgg agcccatcct gggccgcacg 3841 cattacagcc agctgcgcaa gaagagcgga tccgactaca aagaccatga cggtgattat 3901 aaagatcatg acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc 3961 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 4021 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 4081 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 4141 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 4201 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 4261 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 4321 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 4381 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 4441 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 4501 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt 4561 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga 4621 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga 4681 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga 4741 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg 4801 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga 4861 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg 4921 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct 4981 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc 5041 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg 5101 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5161 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5221 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5281 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 5341 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 5401 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 5461 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg 5521 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 5581 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 5641 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 5701 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 5761 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta 5821 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 5881 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 5941 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 6001 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 6061 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 6121 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 6181 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc 6241 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 6301 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg 6361 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 6421 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6481 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6601 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6661 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 6721 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6781 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6841 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6901 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6961 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7021 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7081 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7141 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7201 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7261 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7321 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7381 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7441 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7501 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7561 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7621 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7681 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7741 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7801 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7861 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7921 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7981 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8041 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8101 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8161 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8221 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8281 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8341 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8401 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8461 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8521 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8581 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 8641 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 8701 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 8761 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 8821 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 8881 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 8941 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9001 cagtgccaag ctg //