LOCUS V046968 8899 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046968 VERSION V046968 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..3753 /label="CLK1(NM_004071)" /note="CLK1(NM_004071)" /gene="CLK1" CDS 3760..3825 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3869..4080 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4093..4361 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4388..4987 /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 4994..5582 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5656..5889 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5961..6082 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6101..6236 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6274..6290) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6298..6314 /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(6322..6352) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6367..6388 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6676..7264) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7435..8295) /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(8296..8400) /gene="bla" /label="AmpR promoter" primer_bind 8874..8890 /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 catgagacac tcaaagagaa cttactgtcc tgattgggat 2341 gacaaggatt gggattatgg aaaatggagg agcagcagca gtcataaaag aaggaagaga 2401 tcacatagca gtgcccagga gaacaagcgc tgcaaataca atcactctaa aatgtgtgat 2461 agccattatt tggaaagcag gtctataaat gagaaagatt atcatagtcg acgctacatt 2521 gatgagtaca gaaatgacta cactcaagga tgtgaacctg gacatcgcca aagagaccat 2581 gaaagccggt atcagaacca tagtagcaag tcttctggta gaagtggaag aagtagttat 2641 aaaagcaaac acaggattca ccacagtact tcacatcgtc gttcacatgg gaagagtcac 2701 cgaaggaaaa gaaccaggag tgtagaggat gatgaggagg gtcacctgat ctgtcagagt 2761 ggagacgtac taagtgcaag atatgaaatt gttgatactt taggtgaagg agcttttgga 2821 aaagttgtgg agtgcatcga tcataaagcg ggaggtagac atgtagcagt aaaaatagtt 2881 aaaaatgtgg atagatactg tgaagctgct cgctcagaaa tacaagttct ggaacatctg 2941 aatacaacag accccaacag tactttccgc tgtgtccaga tgttggaatg gtttgagcat 3001 catggtcaca tttgcattgt ttttgaacta ttgggactta gtacttacga cttcattaaa 3061 gaaaatggtt ttctaccatt tcgactggat catatcagaa agatggcata tcagatatgc 3121 aagtctgtga attttttgca cagtaataag ttgactcaca cagacttaaa gcctgaaaac 3181 atcttatttg tgcagtctga ctacacagag gcgtataatc ccaaaataaa acgtgatgaa 3241 cgcaccttaa taaatccaga tattaaagtt gtagactttg gtagtgcaac atatgatgac 3301 gaacatcaca gtacattggt atctacaaga cattatagag cacctgaagt tattttagcc 3361 ctagggtggt cccaaccatg tgatgtctgg agcataggat gcattcttat tgaatactat 3421 cttgggttta ccgtatttcc aacacacgat agtaaggagc atttagcaat gatggaaagg 3481 attcttggac ctctaccaaa acatatgata cagaaaacca ggaaacgtaa atattttcac 3541 cacgatcgat tagactggga tgaacacagt tctgccggca gatatgtttc aagacgctgt 3601 aaacctctga aggaatttat gctttctcaa gatgttgaac atgagcgtct ctttgacctc 3661 attcagaaaa tgttggagta tgatccagcc aaaagaatta ctctcagaga agccttaaag 3721 catcctttct ttgaccttct gaagaaaagt ataggatccg actacaaaga ccatgacggt 3781 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3841 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3901 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3961 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 4021 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 4081 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 4141 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 4201 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 4261 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 4321 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4381 acgctagatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg 4441 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga 4501 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg 4561 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac 4621 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt 4681 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc 4741 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg 4801 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc 4861 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt 4921 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg 4981 tgcctgagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct 5041 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc 5101 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct 5161 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga 5221 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc 5281 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac 5341 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt 5401 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt 5461 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc 5521 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc 5581 gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa 5641 aagaaaaggg gggactggaa gggctaattc actcccaacg aaaataagat ctgctttttg 5701 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag 5761 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc 5821 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa 5881 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat 5941 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca 6001 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 6061 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc 6121 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 6181 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga 6241 cttttgcaga gacggcccaa attcgtaatc atggtcatag ctgtttcctg tgtgaaattg 6301 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 6361 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 6421 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 6481 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 6541 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 6601 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 6661 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 6721 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 6781 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 6841 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 6901 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 6961 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 7021 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 7081 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct 7141 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 7201 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 7261 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 7321 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 7381 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 7441 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 7501 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 7561 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 7621 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 7681 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 7741 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 7801 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 7861 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 7921 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 7981 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 8041 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 8101 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 8161 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 8221 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 8281 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 8341 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 8401 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac 8461 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga 8521 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg 8581 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa 8641 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca 8701 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg 8761 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg 8821 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac 8881 gacggccagt gccaagctg //