LOCUS V040476 9248 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040476 VERSION V040476 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" promoter 2337..3519 /note="strong constitutive promoter for human elongation factor EF-1α" /label="EF-1α promoter" CDS 3548..3904 /label="Rps20(NM_026147)" /note="Rps20(NM_026147)" /gene="Rps20" CDS 3905..3970 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 3998..4584 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 4604..5203 /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 5217..5805 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6012..6645 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6773..6789) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6797..6813 /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(6821..6851) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6866..6887 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7175..7763) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7934..8794) /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(8795..8899) /gene="bla" /label="AmpR promoter" polyA_signal 8947..9081 /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 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg 2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc 2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct 2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtacgaatt 3541 cgccaccatg gcatttaaag ataccggaaa gacgcccgtg gagcccgaag tggcgattca 3601 ccgaattcga atcacgctca ccagccgcaa cgtgaagtcg ctggagaagg tttgtgcgga 3661 cttgatcaga ggcgccaagg aaaagaatct gaaagtgaaa ggaccggtgc gcatgcctac 3721 caagactttg agaatcacta ccagaaaaac accttgtggt gaaggttcca agacctggga 3781 tcgattccag atgaggatcc acaagcgact cattgattta catagtcctt ctgagattgt 3841 taagcagatt acttccatca gtattgagcc gggagttgag gttgaagtca ccattgcaga 3901 tgccgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga 3961 cgatgacaag taggcctgcg atatccagcg gatcccgccc ctctccctcc ccccccccta 4021 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt 4081 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga 4141 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg 4201 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt 4261 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat 4321 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg 4381 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg 4441 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt 4501 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa 4561 cacgatgata agcttgccac aacccacaag gagacgacct tccatgaccg agtacaagcc 4621 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc 4681 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt 4741 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 4801 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 4861 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 4921 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 4981 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 5041 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 5101 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga 5161 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tagacgcgtc tggaacaatc 5221 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 5281 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 5341 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 5401 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 5461 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 5521 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 5581 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 5641 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 5701 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 5761 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg 5821 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc 5881 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca 5941 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 6001 aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg 6061 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt 6121 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt 6181 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat 6241 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca 6301 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa 6361 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg 6421 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag 6481 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat 6541 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact 6601 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc 6661 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt 6721 gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 6781 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 6841 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 6901 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 6961 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 7021 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 7081 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 7141 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 7201 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 7261 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 7321 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 7381 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 7441 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 7501 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 7561 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 7621 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 7681 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 7741 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 7801 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 7861 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 7921 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 7981 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 8041 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 8101 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 8161 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 8221 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 8281 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 8341 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 8401 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 8461 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 8521 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 8581 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 8641 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 8701 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 8761 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 8821 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 8881 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 8941 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 9001 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 9061 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt 9121 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt 9181 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac 9241 ttagtagt //