LOCUS V045414 8398 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045414 VERSION V045414 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /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 1804..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3252 /label="FST(NM_006350)" /note="FST(NM_006350)" /gene="FST" CDS 3259..3324 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3368..3579 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3592..3860 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3887..4486 /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 4493..5081 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5155..5388 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5460..5581 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5600..5735 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5773..5789) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5797..5813 /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(5821..5851) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5866..5887 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6175..6763) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6934..7794) /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(7795..7899) /gene="bla" /label="AmpR promoter" primer_bind 8373..8389 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggtccgc gcgaggcacc agccgggtgg gctttgcctc 2341 ctgctgctgc tgctctgcca gttcatggag gaccgcagtg cccaggctgg gaactgctgg 2401 ctccgtcaag cgaagaacgg ccgctgccag gtcctgtaca agaccgaact gagcaaggag 2461 gagtgctgca gcaccggccg gctgagcacc tcgtggaccg aggaggacgt gaatgacaac 2521 acactcttca agtggatgat tttcaacggg ggcgccccca actgcatccc ctgtaaagaa 2581 acgtgtgaga acgtggactg tggacctggg aaaaaatgcc gaatgaacaa gaagaacaaa 2641 ccccgctgcg tctgcgcccc ggattgttcc aacatcacct ggaagggtcc agtctgcggg 2701 ctggatggga aaacctaccg caatgaatgt gcactcctaa aggcaagatg taaagagcag 2761 ccagaactgg aagtccagta ccaaggcaga tgtaaaaaga cttgtcggga tgttttctgt 2821 ccaggcagct ccacatgtgt ggtggaccag accaataatg cctactgtgt gacctgtaat 2881 cggatttgcc cagagcctgc ttcctctgag caatatctct gtgggaatga tggagtcacc 2941 tactccagtg cctgccacct gagaaaggct acctgcctgc tgggcagatc tattggatta 3001 gcctatgagg gaaagtgtat caaagcaaag tcctgtgaag atatccagtg cactggtggg 3061 aaaaaatgtt tatgggattt caaggttggg agaggccggt gttccctctg tgatgagctg 3121 tgccctgaca gtaagtcgga tgagcctgtc tgtgccagtg acaatgccac ttatgccagc 3181 gagtgtgcca tgaaggaagc tgcctgctcc tcaggtgtgc tactggaagt aaagcactcc 3241 ggatcttgca acggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3301 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3361 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3421 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3481 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3541 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3601 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 3661 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 3721 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 3781 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 3841 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga ccgagtacaa 3901 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc 3961 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg 4021 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg 4081 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg 4141 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc 4201 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct 4261 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct 4321 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc 4381 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc 4441 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg acaatcaacc 4501 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 4561 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 4621 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 4681 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 4741 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 4801 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 4861 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt 4921 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 4981 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 5041 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa 5101 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag 5161 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt 5221 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 5281 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 5341 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat 5401 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga 5461 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 5521 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 5581 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc 5641 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct 5701 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa 5761 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 5821 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 5881 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 5941 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6001 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6061 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6121 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6181 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6241 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 6301 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 6361 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6421 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6481 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 6541 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 6601 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 6661 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 6721 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 6781 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 6841 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 6901 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 6961 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7021 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7081 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 7141 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7201 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 7261 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 7321 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 7381 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 7441 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 7501 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 7561 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 7621 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 7681 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 7741 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 7801 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 7861 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 7921 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 7981 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8041 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 8101 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 8161 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 8221 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 8281 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 8341 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //