LOCUS V023276 7629 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023276 VERSION V023276 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 2301..2483 /label="MT1X(NM_005952)" /note="MT1X(NM_005952)" /gene="MT1X" CDS 2490..2555 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2599..2810 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2823..3091 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3118..3717 /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 3724..4312 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4386..4619 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 4691..4812 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 4831..4966 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5004..5020) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5028..5044 /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(5052..5082) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5097..5118 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5406..5994) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6165..7025) /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(7026..7130) /gene="bla" /label="AmpR promoter" primer_bind 7604..7620 /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta 2281 gagctagcga attcgccacc atggacccca actgctcctg ctcgcctgtt ggctcctgtg 2341 cctgtgccgg ctcctgcaaa tgcaaagagt gcaaatgcac ctcctgcaag aagagctgct 2401 gctcctgctg ccctgtgggc tgtgccaagt gtgcccaggg ctgcatctgc aaagggacgt 2461 cagacaagtg cagctgctgt gccggatccg actacaaaga ccatgacggt gattataaag 2521 atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg atctgcgatc 2581 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg 2641 gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg 2701 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag 2761 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc 2821 gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc 2881 ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct 2941 aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac 3001 ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg 3061 tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagatg 3121 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc 3181 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc 3241 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 3301 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 3361 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 3421 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 3481 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 3541 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 3601 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 3661 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagtc 3721 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 3781 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 3841 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 3901 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 3961 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 4021 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 4081 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg 4141 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 4201 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 4261 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc 4321 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg 4381 gggactggaa gggctaattc actcccaacg aaaataagat ctgctttttg cttgtactgg 4441 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 4501 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 4561 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 4621 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 4681 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 4741 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 4801 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc 4861 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 4921 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 4981 gacggcccaa attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 5041 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 5101 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 5161 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 5221 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 5281 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 5341 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 5401 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 5461 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 5521 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 5581 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 5641 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 5701 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 5761 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 5821 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 5881 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 5941 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 6001 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 6061 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 6121 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 6181 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 6241 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 6301 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 6361 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 6421 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 6481 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 6541 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 6601 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 6661 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 6721 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 6781 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 6841 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 6901 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 6961 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 7021 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 7081 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 7141 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat 7201 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga 7261 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 7321 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca 7381 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta 7441 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg 7501 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg 7561 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt 7621 gccaagctg //