LOCUS V044035 9367 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044035 VERSION V044035 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..4063 /label="EGLN2(NM_053046)" /note="EGLN2(NM_053046)" /gene="EGLN2" CDS 4070..4135 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4155..4654 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4675..5274 /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 5288..5876 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6083..6716 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6845..6861) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6869..6885 /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(6893..6923) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6938..6959 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7247..7835) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8006..8866) /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(8867..8971) /gene="bla" /label="AmpR promoter" polyA_signal 9019..9153 /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 ccatggacag cccgtgccag ccgcagcccc taagtcaggc 2881 tctccctcag ttaccagggt cttcgtcaga gcccttggag cctgagcctg gccgggccag 2941 gatgggagtg gagagttacc tgccctgtcc cctgctcccc tcctaccact gtccaggagt 3001 gcctagtgag gcctcggcag ggagtgggac ccccagagcc acagccacct ctaccactgc 3061 cagccctctt cgggacggtt ttggcgggca ggatggtggt gagctgcggc cgctgcagag 3121 tgaaggcgct gcagcgctgg tcaccaaggg gtgccagcga ttggcagccc agggcgcacg 3181 gcctgaggcc cccaaacgga aatgggccga ggatggtggg gatgcccctt cacccagcaa 3241 acggccctgg gccaggcaag agaaccagga ggcagagcgg gagggtggca tgagctgcag 3301 ctgcagcagt ggcagtggtg aggccagtgc tgggctgatg gaggaggcgc tgccctctgc 3361 gcccgagcgc ctggccctgg actatatcgt gccctgcatg cggtactacg gcatctgcgt 3421 caaggacagc ttcctggggg cagcactggg cggtcgcgtg ctggccgagg tggaggccct 3481 caaacggggt gggcgcctgc gagacgggca gctagtgagc cagagggcga tcccgccgcg 3541 cagcatccgt ggggaccaga ttgcctgggt ggaaggccat gaaccaggct gtcgaagcat 3601 tggtgccctc atggcccatg tggacgccgt catccgccac tgcgcagggc ggctgggcag 3661 ctatgtcatc aacgggcgca ccaaggccat ggtggcgtgt tacccaggca acgggctcgg 3721 gtacgtaagg cacgttgaca atccccacgg cgatgggcgc tgcatcacct gtatctatta 3781 cctgaatcag aactgggacg ttaaggtgca tggcggcctg ctgcagatct tccctgaggg 3841 ccggcccgtg gtagccaaca tcgagccact ctttgaccgg ttgctcattt tctggtctga 3901 ccggcggaac ccccacgagg tgaagccagc ctatgccacc aggtacgcca tcactgtctg 3961 gtattttgat gccaaggagc gggcagcagc caaagacaag tatcagctag catcaggaca 4021 gaaaggtgtc caagtacctg tatcacagcc gcctacgccc accggatccg actacaaaga 4081 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtc 4141 tagataattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt 4201 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac 4261 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac 4321 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca 4381 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa 4441 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca 4501 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg 4561 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc 4621 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc 4681 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 4741 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 4801 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 4861 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 4921 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 4981 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5041 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5101 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5161 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5221 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaccgcgt 5281 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5341 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5401 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5461 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 5521 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 5581 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 5641 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg 5701 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 5761 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 5821 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 5881 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca 5941 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca 6001 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 6061 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc 6121 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca 6181 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag 6241 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc 6301 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta 6361 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga 6421 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact 6481 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc 6541 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 6601 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 6661 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 6721 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 6781 tgagaggcct tgacattgct agcgttttac cgtcgacctc tagctagagc ttggcgtaat 6841 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 6901 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 6961 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 7021 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 7081 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 7141 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 7201 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 7261 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 7321 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 7381 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 7441 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 7501 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 7561 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 7621 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 7681 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 7741 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 7801 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 7861 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 7921 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 7981 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 8041 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 8101 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 8161 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 8221 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 8281 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 8341 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 8401 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 8461 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 8521 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 8581 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 8641 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 8701 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 8761 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 8821 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 8881 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 8941 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 9001 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat 9061 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 9121 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat 9181 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta 9241 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata 9301 tgtactacaa acttagtagt ttttaaagaa attgtatttg ttaaatatgt actacaaact 9361 tagtagt //