LOCUS V044071 9424 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044071 VERSION V044071 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2843..4120 /label="EGLN1(NM_022051)" /note="EGLN1(NM_022051)" /gene="EGLN1" CDS 4127..4192 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4212..4711 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4732..5331 /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 5345..5933 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6140..6773 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6902..6918) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6926..6942 /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(6950..6980) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6995..7016 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7304..7892) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8063..8923) /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(8924..9028) /gene="bla" /label="AmpR promoter" polyA_signal 9076..9210 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 ctcaagcttc gaattcgcca ccatggccaa tgacagcggc gggcccggcg ggccgagccc 2881 gagcgagcga gaccggcagt actgcgagct gtgcgggaag atggagaacc tgctgcgctg 2941 cagccgctgc cgcagctcct tctactgctg caaggagcac cagcgtcagg actggaagaa 3001 gcacaagctc gtgtgccagg gcagcgaggg cgccctcggc cacggagtgg gcccacacca 3061 gcattccggc cccgcgccgc cggctgcagt gccgccgccc agggccgggg cccgggagcc 3121 caggaaggca gcggcgcgcc gggacaacgc ctccggggac gcggccaagg gaaaagtaaa 3181 ggccaagccc ccggccgacc cagcggcggc cgcgtcgccg tgtcgtgcgg ccgccggcgg 3241 ccagggctcg gcggtggctg ccgaagccga gcccggcaag gaggagccgc cggcccgctc 3301 atcgctgttc caggagaagg cgaacctgta ccccccaagc aacacgcccg gggatgcgct 3361 gagccccggc ggcggcctgc ggcccaacgg gcagacgaag cccctgccgg cgctgaagct 3421 ggcgctcgag tacatcgtgc cgtgcatgaa caagcacggc atctgtgtgg tggacgactt 3481 cctcggcaag gagaccggac agcagatcgg cgacgaggtg cgcgccctgc acgacaccgg 3541 gaagttcacg gacgggcagc tggtcagcca gaagagtgac tcgtccaagg acatccgagg 3601 cgataagatc acctggatcg agggcaagga gcccggctgc gaaaccattg ggctgctcat 3661 gagcagcatg gacgacctga tacgccactg taacgggaag ctgggcagct acaaaatcaa 3721 tggccggacg aaagccatgg ttgcttgtta tccgggcaat ggaacgggtt atgtacgtca 3781 tgttgataat ccaaatggag atggaagatg tgtgacatgt atatattatc ttaataaaga 3841 ctgggatgcc aaggtaagtg gaggtatact tcgaattttt ccagaaggca aagcccagtt 3901 tgctgacatt gaacccaaat ttgatagact gctgtttttc tggtctgacc gtcgcaaccc 3961 tcatgaagta caaccagcat atgctacaag gtacgcaata actgtttggt attttgatgc 4021 agatgagaga gcacgagcta aagtaaaata tctaacaggt gaaaaaggtg tgagggttga 4081 actcaataaa ccttcagatt cggtcggtaa agacgtcttc ggatccgact acaaagacca 4141 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtctag 4201 ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc 4261 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc 4321 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc 4381 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat 4441 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg 4501 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag 4561 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc 4621 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct 4681 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag 4741 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4801 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4861 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4921 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4981 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 5041 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 5101 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 5161 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 5221 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5281 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg 5341 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 5401 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 5461 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5521 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5581 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5641 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5701 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct 5761 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5821 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5881 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat 5941 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct 6001 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc 6061 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 6121 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt 6181 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg 6241 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca 6301 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg 6361 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca 6421 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct 6481 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg 6541 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct 6601 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 6661 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 6721 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag 6781 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga 6841 gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg gcgtaatcat 6901 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 6961 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 7021 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 7081 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 7141 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 7201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 7261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 7321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 7381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 7441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 7501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 7561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 7621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 7681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 7741 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 7801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 7861 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 7921 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 7981 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 8041 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 8101 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 8161 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 8221 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 8281 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 8341 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 8401 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 8461 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 8521 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 8581 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 8641 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 8701 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 8761 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 8821 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 8881 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 8941 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 9001 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 9061 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc 9121 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 9181 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat 9241 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc 9301 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt 9361 actacaaact tagtagtttt taaagaaatt gtatttgtta aatatgtact acaaacttag 9421 tagt //