LOCUS V046000 9284 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046000 VERSION V046000 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 2058..2175 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" enhancer 2227..2530 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2531..2734 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2826..2915 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" CDS 2931..4445 /label="PFKFB2(NM_006212)" /note="PFKFB2(NM_006212)" /gene="PFKFB2" CDS 4452..4523 /codon_start=1 /product="three tandem FLAG® epitope tags" /transl_table=1 /translation="DYKDDDDKDYKDDDDKDYKDDDDK" /label="3xFLAG" promoter 4541..5040 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5061..5660 /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 5674..6262 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6337..6570 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" promoter 6775..6879 /gene="bla" /label="AmpR promoter" CDS 6880..7740 /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" rep_origin 7911..8499 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" protein_bind 8910..8931 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 8946..8976 /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8984..9000 /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" primer_bind 9008..9024 /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" 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 tatcgccgaa ttcacaaatg 2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa 2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca 2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtgga 2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 2701 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg 2761 gagacgccat ccacgctgtt ttgacctcca tagaagacac cggcggccac gcgtgcggcc 2821 gcatggagca gaaactcatc tctgaagaag atctggaaca aaagttgatt tcagaagaag 2881 atctggaaca gaagctcatc tctgaggaag atctggcccg ggcgggatcc atgtctgggg 2941 catcttcctc agaacagaac aacaacagct atgaaaccaa aaccccaaat cttcgaatgt 3001 cagagaagaa atgttcatgg gcctcctaca tgaccaactc cccgactctg atcgttatga 3061 ttggtttgcc agcccggggt aaaacctacg tgtccaagaa actaacacgc tacctcaact 3121 ggattggagt ccccaccaaa gtgtttaatc ttggggtgta tcggcgtgaa gcagtcaagt 3181 cctataagtc ctacgacttc tttcggcatg acaatgagga ggccatgaag atccgcaaac 3241 agtgtgctct ggtggcgctg gaagatgtta aggcgtatct cactgaggag aatggtcaga 3301 ttgcggtgtt tgatgccacc aatacaaccc gggagaggag ggacatgatt ttgaactttg 3361 ctgaacagaa ttccttcaag gtattctttg tggaatccgt ctgtgatgat cctgatgtca 3421 ttgctgccaa tattctggag gttaaggtat caagccctga ctatcctgaa aggaacagag 3481 agaacgtgat ggaggacttc ctgaagagaa ttgaatgcta caaagttacc taccgacctc 3541 ttgacccaga caactatgac aaggatcttt ctttcatcaa ggtgataaac gtgggccagc 3601 gatttttagt caacagagtc caggactaca tccagagcaa gatagtctac tacctcatga 3661 atatccacgt ccagcctcgc accatttacc tttgccggca tggagaaagc gagttcaatc 3721 tcttggggaa gattgggggt gactctggcc tctcggtgcg gggaaagcag tttgcccaag 3781 ctctaaggaa atttctggag gaacaggaaa taacagacct caaagtgtgg acaagccagt 3841 tgaagaggac catacagact gctgaatctc tcggggtgcc ctatgagcag tggaagattc 3901 tgaatgagat tgatgctggt gtgtgtgaag agatgaccta tgcagagatt gagaaacggt 3961 acccagaaga gtttgcactt cgagatcaag agaagtatct gtatcgatat cctggtgggg 4021 agtcatacca ggacctggtg cagcggctgg agcctgtcat catggagctg gaacgtcagg 4081 gcaatgtcct cgtcatctcc caccaggctg tcatgcgctg cctcctggcc tacttcttgg 4141 ataagggcgc agatgagcta ccatacttga gatgccctct ccataccatc ttcaaactta 4201 ctcctgtggc ctatgggtgc aaagtggaaa caattaaact taacgtggag gctgtgaaca 4261 cgcaccgtga caagccaact aacaacttcc ccaagaacca aacccctgta aggatgagaa 4321 ggaacagctt tacgcctctg tccagttcga atacaataag gcgtccaaga aattacagtg 4381 ttgggagccg gcccctcaag cccctcagcc ctctccgtgc ccaggacatg caagaagggg 4441 ccgacctcga ggattacaag gatgacgacg ataaggacta taaggacgat gatgacaagg 4501 actacaaaga tgatgacgat aaatagtcga gaattctacc gggtagggga ggcgcttttc 4561 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt 4621 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt 4681 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg 4741 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag 4801 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag 4861 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc 4921 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc 4981 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg 5041 acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg 5101 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc 5161 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc 5221 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg 5281 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc 5341 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg 5401 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc 5461 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg 5521 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc 5581 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga 5641 cccgcaagcc cggtgcctga atcgatagat cctaatcaac ctctggatta caaaatttgt 5701 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct 5761 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat 5821 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg 5881 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag 5941 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc 6001 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg 6061 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc 6121 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc 6181 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc 6241 tccctttggg ccgcctcccc gcctgagatc ctttaagacc aatgacttac aaggcagctg 6301 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac 6361 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct 6421 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag 6481 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac 6541 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt 6601 atttataact tgcaaagaaa tgaatatcag agagtgagag gcccgggtta attaaggaaa 6661 gggctagatc attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat 6721 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga 6781 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 6841 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 6901 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 6961 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 7021 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 7081 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag 7141 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 7201 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 7261 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 7321 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 7381 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 7441 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 7501 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 7561 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 7621 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 7681 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 7741 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 7801 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 7861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 7921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 7981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 8041 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 8101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 8161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 8221 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 8281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 8341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 8401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 8461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 8521 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 8581 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 8641 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg 8701 cctctccccg cgcgttggcc gattcattaa tgcagcaagc tcatggctga ctaatttttt 8761 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 8821 gcttttttgg aggcctaggc ttttgcaaaa agctccccgt ggcacgacag gtttcccgac 8881 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc 8941 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa 9001 tttcacacag gaaacagcta tgacatgatt acgaatttca caaataaagc atttttttca 9061 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac 9121 tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact 9181 gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt 9241 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt //