LOCUS V024462 9908 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024462 VERSION V024462 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 3551..3568 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" CDS 3569..4567 /label="XCR1(NM_001381860)" /note="XCR1(NM_001381860)" /gene="XCR1" CDS 4574..4639 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4658..5244 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5264..5863 /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 5877..6465 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6672..7305 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7433..7449) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7457..7473 /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(7481..7511) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7526..7547 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7835..8423) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8594..9454) /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(9455..9559) /gene="bla" /label="AmpR promoter" polyA_signal 9607..9741 /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 caccatcacc atcaccatat ggagtcctca ggcaacccag agagcaccac 3601 ctttttttac tatgaccttc agagccagcc gtgtgagaac caggcctggg tctttgctac 3661 cctcgccacc actgtcctat actgcctggt gtttctcctc agcctagtgg gcaacagcct 3721 ggtcctgtgg gtcctggtga agtatgagag cctggagtcc ctcaccaaca tcttcatcct 3781 caacctgtgc ctctcagacc tggtgttcgc ctgcttgttg cctgtgtgga tctccccata 3841 ccactggggc tgggtgctgg gagacttcct ctgcaaactc ctcaatatga tcttctccat 3901 cagcctctac agcagcatct tcttcctgac catcatgacc atccaccgct acctgtcggt 3961 agtgagcccc ctctccaccc tgcgcgtccc caccctccgc tgccgggtgc tggtgaccat 4021 ggctgtgtgg gtagccagca tcctgtcctc catcctcgac accatcttcc acaaggtgct 4081 ttcttcgggc tgtgattatt ccgaactcac gtggtacctc acctccgtct accagcacaa 4141 cctcttcttc ctgctgtccc tggggattat cctgttctgc tacgtggaga tcctcaggac 4201 cctgttccgc tcacgctcca agcggcgcca ccgcacggtc aagctcatct tcgccatcgt 4261 ggtggcctac ttcctcagct ggggtcccta caacttcacc ctgtttctgc agacgctgtt 4321 tcggacccag atcatccgga gctgcgaggc caaacagcag ctagaatacg ccctgctcat 4381 ctgccgcaac ctcgccttct cccactgctg ctttaacccg gtgctctatg tcttcgtggg 4441 ggtcaagttc cgcacacacc tgaaacatgt tctccggcag ttctggttct gccggctgca 4501 ggcacccagc ccagcctcga tcccccactc ccctggtgcc ttcgcctatg agggcgcctc 4561 cttctactct agagactaca aagaccatga cggtgattat aaagatcatg acatcgacta 4621 caaggatgac gatgacaagt agcggccgcg gatcccgccc ctctccctcc ccccccccta 4681 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt 4741 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga 4801 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg 4861 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt 4921 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat 4981 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg 5041 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg 5101 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt 5161 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa 5221 cacgatgata agcttgccac aacccacaag gagacgacct tccatgaccg agtacaagcc 5281 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc 5341 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt 5401 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 5461 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 5521 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 5581 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 5641 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 5701 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 5761 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga 5821 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tagacgcgtc tggaacaatc 5881 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 5941 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 6001 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 6061 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 6121 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 6181 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 6241 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 6301 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 6361 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 6421 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg 6481 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc 6541 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca 6601 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 6661 aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg 6721 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt 6781 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt 6841 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat 6901 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca 6961 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa 7021 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg 7081 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag 7141 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat 7201 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact 7261 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc 7321 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt 7381 gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 7441 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 7501 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 7561 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 7621 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 7681 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 7741 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 7801 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 7861 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 7921 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 7981 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 8041 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 8101 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 8161 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 8221 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 8281 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 8341 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 8401 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 8461 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 8521 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 8581 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 8641 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 8701 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 8761 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 8821 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 8881 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 8941 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 9001 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 9061 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 9121 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 9181 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 9241 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 9301 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 9361 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 9421 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 9481 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 9541 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 9601 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 9661 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 9721 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt 9781 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt 9841 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac 9901 ttagtagt //