LOCUS V028203 9667 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028203 VERSION V028203 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..4521 /label="DTX3L(NM_138287)" /note="DTX3L(NM_138287)" /gene="DTX3L" CDS 4528..4593 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4637..4848 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4861..5129 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5156..5755 /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 5762..6350 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6424..6657 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6729..6850 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6869..7004 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7042..7058) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7066..7082 /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(7090..7120) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7135..7156 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7444..8032) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8203..9063) /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(9064..9168) /gene="bla" /label="AmpR promoter" primer_bind 9642..9658 /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 catggcctcc cacctgcgcc cgccgtcccc gctcctcgtg 2341 cgggtgtaca agtccggccc ccgagtacga aggaagctgg agagctactt ccagagctct 2401 aagtcctcgg gcggcgggga gtgcacggtc agcacccagg aacacgaagc cccgggcacc 2461 ttccgggtgg agttcagtga aagggcagct aaggagagag tgttgaaaaa aggagagcac 2521 caaatacttg ttgacgaaaa acctgtgccc attttcctgg tacccactga aaattcaata 2581 aagaagaaca cgagacctca aatttcttca ctgacacaat cacaagcaga aacaccgtct 2641 ggtgatatgc atcaacatga aggacatatt cctaatgctg tggattcctg tctccaaaag 2701 atctttctta ctgtaacagc tgacctgaac tgtaacctgt tctccaaaga gcagagggca 2761 tacataacca cactgtgccc tagtatcaga aaaatggaag gtcacgatgg aattgagaag 2821 gtgtgtggtg acttccaaga cattgaaaga atacatcaat ttttgagtga gcagttcctg 2881 gaaagtgagc agaaacaaca attttcccct tcaatgacag agaggaagcc actcagtcag 2941 caggagaggg acagctgcat ttctccttct gaaccagaaa ccaaggcaga acaaaaaagc 3001 aactattttg aagttccctt gccttacttt gaatacttta aatatatctg tcctgataaa 3061 atcaactcaa tagagaaaag atttggtgta aacattgaaa tccaggagag ttctccaaat 3121 atggtctgtt tagatttcac ctcaagtcga tcaggtgacc tggaagcagc tcgtgagtct 3181 tttgctagtg aatttcagaa gaacacagaa cctctgaagc aagaatgtgt ctctttagca 3241 gacagtaagc aggcaaataa attcaaacag gaattgaatc accagtttac aaagctcctt 3301 ataaaggaga aaggaggcga attaactctc cttgggaccc aagatgacat ttcagctgcc 3361 aaacaaaaaa tctctgaagc ttttgtcaag atacctgtga aactatttgc tgccaattac 3421 atgatgaatg taattgaggt tgatagtgcc cactataaac ttttagaaac tgaattacta 3481 caggagatat cagagatcga aaaaaggtat gacatttgca gcaaggtttc tgagaaaggt 3541 cagaaaacct gcattctgtt tgaatccaag gacaggcagg tagatctatc tgtgcatgct 3601 tatgcaagtt tcatcgatgc ctttcaacat gcctcatgtc agttgatgag agaagttctt 3661 ttactgaagt ctttgggcaa ggagagaaag cacttacatc agaccaagtt tgctgatgac 3721 tttagaaaaa gacatccaaa tgtacacttt gtgctaaatc aagagtcaat gactttgact 3781 ggtttgccaa atcaccttgc aaaggcgaag cagtatgttc taaaaggagg aggaatgtct 3841 tcattggctg gaaagaaatt gaaagagggt catgaaacac cgatggacat tgatagcgat 3901 gattccaaag cagcttctcc gccactcaag ggctctgtga gttctgaggc ctcagaactg 3961 gacaagaagg aaaagggcat ctgtgtcatc tgtatggaca ccattagtaa caaaaaagtg 4021 ctaccaaagt gcaagcatga attctgcgcc ccttgtatca acaaagccat gtcatataag 4081 ccaatctgtc ccacatgcca gacttcctat ggtattcaga aaggaaatca gccagaggga 4141 agcatggttt tcactgtttc aagagactca cttccaggtt atgagtcctt tggcaccatt 4201 gtgattactt attctatgaa agcaggcata caaacagaag aacacccaaa cccaggaaag 4261 agataccctg gaatacagcg aactgcatac ttgcctgata ataaggaagg aaggaaggtt 4321 ttgaaactgc tttatagggc ctttgaccaa aagctgattt ttacagtggg gtactctcgc 4381 gtattaggag tctcagatgt catcacttgg aatgatattc accacaaaac atcccggttt 4441 ggaggaccag aaatgtatgg ctatcctgat ccttcttacc tgaaacgtgt caaagaggag 4501 ctgaaagcca aaggaattga gggatccgac tacaaagacc atgacggtga ttataaagat 4561 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 4621 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 4681 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 4741 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 4801 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 4861 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 4921 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 4981 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 5041 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 5101 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 5161 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 5221 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 5281 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 5341 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 5401 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 5461 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 5521 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 5581 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 5641 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 5701 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 5761 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5821 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5881 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5941 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 6001 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 6061 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 6121 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 6181 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 6241 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 6301 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 6361 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 6421 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 6481 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 6541 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 6601 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 6661 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6721 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6781 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6841 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6901 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6961 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 7021 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 7081 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 7141 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7201 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7261 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7321 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7381 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7441 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7501 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7561 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7621 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7681 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7741 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7801 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7861 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7921 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7981 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 8041 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 8101 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 8161 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8221 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8281 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8341 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8401 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8461 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8521 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8581 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8641 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8701 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8761 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8821 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8881 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8941 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 9001 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 9061 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 9121 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9181 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9241 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9301 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9361 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9421 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9481 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9541 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9601 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9661 caagctg //