LOCUS V020279 7933 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020279 VERSION V020279 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2289..2855 /codon_start=1 /gene="" /product="human small GTPase proto-oncoprotein" /protein_id="" /transl_table=1 /translation="MTEYKLVVVGAGDVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI RKHKEKMSKDGKKKKKKSKTKCVIM*" /label="K-Ras" promoter 2903..3114 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3127..3395 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3422..4021 /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 4028..4616 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4690..4923 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 4995..5116 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5135..5270 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5308..5324) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5332..5348 /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(5356..5386) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5401..5422 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5710..6298) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6469..7329) /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(7330..7434) /gene="bla" /label="AmpR promoter" primer_bind 7908..7924 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagccaccat gactgaatat aaacttgtgg tagttggagc tggtgacgta ggcaagagtg 2341 ccttgacgat acagctaatt cagaatcatt ttgtggacga atatgatcca acaatagagg 2401 attcctacag gaagcaagta gtaattgatg gagaaacctg tctcttggat attctcgaca 2461 cagcaggtca agaggagtac agtgcaatga gggaccagta catgaggact ggggagggct 2521 ttctttgtgt atttgccata aataatacta aatcatttga agatattcac cattatagag 2581 aacaaattaa aagagttaag gactctgaag atgtacctat ggtcctagta ggaaataaat 2641 gtgatttgcc ttctagaaca gtagacacaa aacaggctca ggacttagca agaagttatg 2701 gaattccttt tattgaaaca tcagcaaaga caagacaggg tgttgatgat gccttctata 2761 cattagttcg agaaattcga aaacataaag aaaagatgag caaagatggt aaaaagaaga 2821 aaaagaagtc aaagacaaag tgtgtaatta tgtaaggatc cgcggccgcg aaggatctgc 2881 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 2941 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 3001 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 3061 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 3121 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 3181 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 3241 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 3301 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 3361 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 3421 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt 3481 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga 3541 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga 3601 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga 3661 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg 3721 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga 3781 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg 3841 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct 3901 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc 3961 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg 4021 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 4081 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 4141 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 4201 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 4261 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 4321 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 4381 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg 4441 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 4501 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 4561 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 4621 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 4681 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta 4741 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 4801 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 4861 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 4921 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 4981 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 5041 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 5101 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc 5161 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 5221 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg 5281 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 5341 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 5401 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 5461 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 5521 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 5581 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 5641 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 5701 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 5761 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 5821 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 5881 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 5941 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 6001 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 6061 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 6121 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 6181 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 6241 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 6301 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 6361 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 6421 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 6481 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 6541 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 6601 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6661 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6721 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 6781 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6841 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6901 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6961 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 7021 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 7081 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 7141 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 7201 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 7261 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 7321 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 7381 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 7441 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 7501 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 7561 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 7621 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 7681 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 7741 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 7801 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 7861 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 7921 cagtgccaag ctg //