LOCUS V044424 8704 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044424 VERSION V044424 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..3534 /label="PDK4(NM_002612)" /note="PDK4(NM_002612)" /gene="PDK4" CDS 3541..3630 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" promoter 3674..3885 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3898..4166 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4193..4792 /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 4799..5387 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5461..5694 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5766..5887 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5906..6041 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6079..6095) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6103..6119 /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(6127..6157) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6172..6193 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6481..7069) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7240..8100) /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(8101..8205) /gene="bla" /label="AmpR promoter" primer_bind 8679..8695 /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 catgaaggcg gcccgcttcg tgctgcgcag cgctggctcg 2341 ctcaacggcg ccggcctggt gccccgagag gtggagcatt tctcgcgcta cagcccgtcc 2401 ccgctgtcca tgaagcagct actggacttt ggttcagaaa atgcatgtga aagaacttct 2461 tttgcatttt tgcgacaaga attgcctgtg agactcgcca acattctgaa ggaaattgat 2521 atcctcccga cccaattagt aaatacctct tcagtgcaat tggttaaaag ctggtatata 2581 cagagcctga tggatttggt ggaattccat gagaaaagcc cagatgacca gaaagcatta 2641 tcagactttg tagatacact catcaaagtt cgaaatagac accataatgt agtccctaca 2701 atggcacaag gaatcataga gtataaagat gcctgtacag ttgacccagt caccaatcaa 2761 aatcttcaat atttcttgga tcgattttac atgaaccgta tttctactcg gatgctgatg 2821 aaccagcaca ttcttatatt tagtgactca cagacaggaa acccaagcca cattggaagc 2881 attgatccta actgtgatgt ggtagcagtg gtccaagatg cctttgagtg ttcaaggatg 2941 ctctgtgatc agtattattt atcatctcca gaattaaagc ttacacaagt gaatggaaaa 3001 tttccagacc aaccaattca catcgtgtat gttccttctc acctccatca tatgctcttt 3061 gaactattta agaatgcaat gcgggcaaca gttgaacacc aggaaaatca gccttccctt 3121 acaccaatag aggttattgt tgtcttggga aaagaagacc ttaccattaa gatttcagac 3181 agaggaggtg gtgttcccct gagaattatt gaccgcctct ttagttatac atactccact 3241 gcaccaacgc ctgtgatgga taattcccgg aatgctcctt tggctggttt tggttacggc 3301 ttgccaattt ctcgtctgta tgcaaagtac tttcaaggag atctgaatct ctactcttta 3361 tcaggatatg gaacagatgc tatcatctac ttaaaggctt tgtcttctga gtctatagaa 3421 aaacttccag tttttaacaa gtcagccttc aaacattatc agatgagctc tgaggctgat 3481 gactggtgta tcccaagcag ggaaccaaag aacctggcaa aagaagtggc catgggatcc 3541 gaacaaaaac taatatcaga ggaagatttg gaacagaaac taatcagtga agaagattta 3601 gagcaaaagt taatttccga ggaggactta tagcggccgc gaaggatctg cgatcgctcc 3661 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 3721 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 3781 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 3841 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 3901 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 3961 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 4021 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 4081 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 4141 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 4201 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 4261 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 4321 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4381 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4441 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4501 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4561 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 4621 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 4681 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 4741 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 4801 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 4861 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 4921 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 4981 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5041 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5101 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5161 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 5221 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5281 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5341 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 5401 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5461 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5521 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5581 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 5641 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 5701 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 5761 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 5821 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 5881 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 5941 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6001 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6061 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6121 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6181 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6241 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6901 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 6961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7141 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7201 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7261 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7321 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7381 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7441 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7501 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7561 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7621 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7681 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7741 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7801 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7861 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7921 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7981 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8041 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8101 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8161 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8221 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8281 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8341 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8401 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8461 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8521 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8581 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8641 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8701 gctg //