LOCUS V030745 10028 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030745 VERSION V030745 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 2303..4930 /label="KPNB1(NM_002265)" /note="KPNB1(NM_002265)" /gene="KPNB1" CDS 4937..4954 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" promoter 4998..5209 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 5222..5490 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5517..6116 /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 6123..6711 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6785..7018 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7090..7211 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7230..7365 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7403..7419) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7427..7443 /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(7451..7481) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7496..7517 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7805..8393) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8564..9424) /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(9425..9529) /gene="bla" /label="AmpR promoter" primer_bind 10003..10019 /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 aattcggcca ccatggagct gatcaccatt ctcgagaaga ccgtgtctcc 2341 cgatcggctg gagctggaag cggcgcagaa gttcctggag cgtgcggccg tggagaacct 2401 gcccactttc cttgtggaac tgtccagagt gctggcaaat ccaggaaaca gtcaggttgc 2461 cagagttgca gctggtctac aaatcaagaa ctctttgaca tctaaagatc cagatatcaa 2521 ggcacaatat cagcagaggt ggcttgctat tgatgctaat gctcgacgag aagtcaagaa 2581 ctatgttttg cagacattgg gtacagaaac ttaccggcct agttctgcct cacagtgtgt 2641 ggctggtatt gcttgtgcag agatcccagt aaaccagtgg ccagaactca ttcctcagct 2701 ggtggccaat gtcacaaacc ccaacagcac agagcacatg aaggagtcga cattggaagc 2761 catcggttat atttgccaag atatagaccc agagcagcta caagataaat ccaatgagat 2821 tctgactgcc ataatccagg ggatgaggaa agaagagcct agtaataatg tgaagctagc 2881 tgctacgaat gcactcctga actcattgga gttcaccaaa gcaaactttg ataaagagtc 2941 tgaaaggcac tttattatgc aggtggtctg tgaagccaca cagtgtccag atacgagggt 3001 acgagtggct gctttacaga atctggtgaa gataatgtcc ttatattatc agtacatgga 3061 gacatatatg ggtcctgctc tttttgcaat cacaatcgaa gcaatgaaaa gtgacattga 3121 tgaggtggct ttacaaggga tagaattctg gtccaatgtc tgtgatgagg aaatggattt 3181 ggccattgaa gcttcagagg cagcagaaca aggacggccc cctgagcaca ccagcaagtt 3241 ttatgcgaag ggagcactac agtatctggt tccaatcctc acacagacac taactaaaca 3301 ggacgaaaat gatgatgacg atgactggaa cccctgcaaa gcagcagggg tgtgcctcat 3361 gcttctggcc acctgctgtg aagatgacat tgtcccacat gtcctcccct tcattaaaga 3421 acacatcaag aacccagatt ggcggtaccg ggatgcagca gtgatggctt ttggttgtat 3481 cttggaagga ccagagccca gtcagctcaa accactagtt atacaggcta tgcccaccct 3541 aatagaatta atgaaagacc ccagtgtagt tgttcgagat acagctgcat ggactgtagg 3601 cagaatttgt gagctgcttc ctgaagctgc catcaatgat gtctacttgg ctcccctgct 3661 acagtgtctg attgagggtc tcagtgctga acccagagtg gcttcaaatg tgtgctgggc 3721 tttctccagt ctggctgaag ctgcttatga agctgcagac gttgctgatg atcaggaaga 3781 accagctact tactgcttat cttcttcatt tgaactcata gttcagaagc tcctagagac 3841 tacagacaga cctgatggac accagaacaa cctgaggagt tctgcatatg aatctctgat 3901 ggaaattgtg aaaaacagtg ccaaggattg ttatcctgct gtccagaaaa cgactttggt 3961 catcatggaa cgactgcaac aggttcttca gatggagtca catatccaga gcacatccga 4021 tagaatccag ttcaatgacc ttcagtcttt actctgtgca actcttcaga atgttcttcg 4081 gaaagtgcaa catcaagatg ctttgcagat ctctgatgtg gttatggcct ccctgttaag 4141 gatgttccaa agcacagctg ggtctggggg agtacaagag gatgccctga tggcagttag 4201 cacactggtg gaagtgttgg gtggtgaatt cctcaagtac atggaggcct ttaaaccctt 4261 cctgggcatt ggattaaaaa attatgctga ataccaggtt tgtttggcag ctgtgggctt 4321 agtgggagac ttgtgccgtg ccctgcaatc caacatcata cctttctgtg acgaggtgat 4381 gcagctgctt ctggaaaatt tggggaatga gaacgtccac aggtctgtga agccgcagat 4441 tctgtcagtg tttggtgata ttgcccttgc tattggagga gagtttaaaa aatacttaga 4501 ggttgtattg aatactcttc agcaggcctc ccaagcccag gtggacaagt cagactatga 4561 catggtggat tatctgaatg agctaaggga aagctgcttg gaagcctata ctggaatcgt 4621 ccagggatta aagggggatc aggagaacgt acacccggat gtgatgctgg tacaacccag 4681 agtagaattt attctgtctt tcattgacca cattgctgga gatgaggatc acacagatgg 4741 agtagtagct tgtgctgctg gactaatagg ggacttatgt acagcatttg ggaaggatgt 4801 actgaaatta gtagaagcta ggccaatgat ccatgaattg ttaactgaag ggcggagatc 4861 gaagactaac aaagcaaaaa cccttgctac atgggcaaca aaagaactga ggaaactgaa 4921 gaaccaagct ggatcccatc accatcacca tcactagcgg ccgcgaagga tctgcgatcg 4981 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg 5041 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga 5101 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt 5161 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg 5221 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg 5281 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta 5341 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc 5401 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt 5461 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga 5521 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca 5581 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc 5641 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 5701 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 5761 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 5821 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 5881 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 5941 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 6001 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 6061 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg 6121 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 6181 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 6241 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 6301 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 6361 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 6421 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 6481 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc 6541 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 6601 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 6661 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct 6721 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 6781 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg 6841 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6901 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6961 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 7021 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 7081 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 7141 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 7201 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 7261 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 7321 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 7381 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 7441 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 7501 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 7561 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 7621 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 7681 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 7741 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 7801 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 7861 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 7921 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 7981 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 8041 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 8101 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 8161 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 8221 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 8281 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 8341 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 8401 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 8461 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 8521 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 8581 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 8641 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 8701 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 8761 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 8821 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 8881 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8941 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 9001 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 9061 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 9121 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 9181 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 9241 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 9301 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 9361 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 9421 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 9481 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 9541 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 9601 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 9661 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 9721 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 9781 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 9841 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 9901 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 9961 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 10021 ccaagctg //