LOCUS V033978 8830 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033978 VERSION V033978 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..3684 /label="TNFRSF1B(NM_001066)" /note="TNFRSF1B(NM_001066)" /gene="TNFRSF1B" CDS 3691..3756 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3800..4011 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4024..4292 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4319..4918 /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 4925..5513 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5587..5820 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5892..6013 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6032..6167 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6205..6221) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6229..6245 /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(6253..6283) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6298..6319 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6607..7195) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7366..8226) /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(8227..8331) /gene="bla" /label="AmpR promoter" primer_bind 8805..8821 /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 catggcgccc gtcgccgtct gggccgcgct ggccgtcgga 2341 ctggagctct gggctgcggc gcacgccttg cccgcccagg tggcatttac accctacgcc 2401 ccggagcccg ggagcacatg ccggctcaga gaatactatg accagacagc tcagatgtgc 2461 tgcagcaaat gctcgccggg ccaacatgca aaagtcttct gtaccaagac ctcggacacc 2521 gtgtgtgact cctgtgagga cagcacatac acccagctct ggaactgggt tcccgagtgc 2581 ttgagctgtg gctcccgctg tagctctgac caggtggaaa ctcaagcctg cactcgggaa 2641 cagaaccgca tctgcacctg caggcccggc tggtactgcg cgctgagcaa gcaggagggg 2701 tgccggctgt gcgcgccgct gcgcaagtgc cgcccgggct tcggcgtggc cagaccagga 2761 actgaaacat cagacgtggt gtgcaagccc tgtgccccgg ggacgttctc caacacgact 2821 tcatccacgg atatttgcag gccccaccag atctgtaacg tggtggccat ccctgggaat 2881 gcaagcatgg atgcagtctg cacgtccacg tcccccaccc ggagtatggc cccaggggca 2941 gtacacttac cccagccagt gtccacacga tcccaacaca cgcagccaac tccagaaccc 3001 agcactgctc caagcacctc cttcctgctc ccaatgggcc ccagcccccc agctgaaggg 3061 agcactggcg acttcgctct tccagttgga ctgattgtgg gtgtgacagc cttgggtcta 3121 ctaataatag gagtggtgaa ctgtgtcatc atgacccagg tgaaaaagaa gcccttgtgc 3181 ctgcagagag aagccaaggt gcctcacttg cctgccgata aggcccgggg tacacagggc 3241 cccgagcagc agcacctgct gatcacagcg ccgagctcca gcagcagctc cctggagagc 3301 tcggccagtg cgttggacag aagggcgccc actcggaacc agccacaggc accaggcgtg 3361 gaggccagtg gggccgggga ggcccgggcc agcaccggga gctcagattc ttcccctggt 3421 ggccatggga cccaggtcaa tgtcacctgc atcgtgaacg tctgtagcag ctctgaccac 3481 agctcacagt gctcctccca agccagctcc acaatgggag acacagattc cagcccctcg 3541 gagtccccga aggacgagca ggtccccttc tccaaggagg aatgtgcctt tcggtcacag 3601 ctggagacgc cagagaccct gctggggagc accgaagaga agcccctgcc ccttggagtg 3661 cctgatgctg ggatgaagcc cagtggatcc gactacaaag accatgacgg tgattataaa 3721 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3781 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3841 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3901 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3961 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 4021 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 4081 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4141 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4201 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4261 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat 4321 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg 4381 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg 4441 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 4501 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 4561 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 4621 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 4681 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 4741 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 4801 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 4861 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt 4921 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4981 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 5041 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5101 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5161 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5221 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5281 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 5341 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5401 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5461 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 5521 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 5581 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 5641 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 5701 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 5761 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 5821 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 5881 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5941 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 6001 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 6061 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 6121 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 6181 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6241 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6301 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6361 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6421 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6481 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6541 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6601 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6661 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6721 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6781 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6841 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6901 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 6961 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7021 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7081 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7141 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7201 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7261 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7321 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7381 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7441 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7501 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7561 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7621 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7681 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7741 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7801 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7861 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7921 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 7981 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8041 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8101 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8161 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8221 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8281 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8341 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8401 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8461 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8521 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8581 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8641 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8701 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8761 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 8821 tgccaagctg //