LOCUS V038721 9824 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038721 VERSION V038721 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..2946 /label="HMGB1(NM_001313893)" /note="HMGB1(NM_001313893)" /gene="HMGB1" promoter 2996..3207 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3220..3488 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3517..3526 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3547..5196 /EC_number="" /codon_start=1 /gene="luc+" /note="enhanced luc+ version of the luciferase gene" /product="firefly luciferase" /protein_id="" /transl_table=1 /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" /label="luciferase" CDS 5266..5319 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 5320..5919 /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 5920..6507 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6581..6814 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6886..7007 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7026..7161 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7199..7215) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7223..7239 /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(7247..7277) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7292..7313 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7601..8189) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8360..9220) /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(9221..9325) /gene="bla" /label="AmpR promoter" primer_bind 9799..9815 /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 gggaggtcta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgggcaaa ggagatccta agaagccgag aggcaaaatg 2341 tcatcatatg cattttttgt gcaaacttgt cgggaggagc ataagaagaa gcacccagat 2401 gcttcagtca acttctcaga gttttctaag aagtgctcag agaggtggaa gaccatgtct 2461 gctaaagaga aaggaaaatt tgaagatatg gcaaaagcgg acaaggcccg ttatgaaaga 2521 gaaatgaaaa cctatatccc tcccaaaggg gagacaaaaa agaagttcaa ggatcccaat 2581 gcacccaaga ggcctccttc ggccttcttc ctcttctgct ctgagtatcg cccaaaaatc 2641 aaaggagaac atcctggcct gtccattggt gatgttgcga agaaactggg agagatgtgg 2701 aataacactg ctgcagatga caagcagcct tatgaaaaga aggctgcgaa gctgaaggaa 2761 aaatacgaaa aggatattgc tgcatatcga gctaaaggaa agcctgatgc agcaaaaaag 2821 ggagttgtca aggctgaaaa aagcaagaaa aagaaggaag aggaggaaga tgaggaagat 2881 gaagaggatg aggaggagga ggaagatgaa gaagatgaag atgaagaaga agatgatgat 2941 gatgaataag gatccgcggc cgcaaggatc tgcgatcgct ccggtgcccg tcagtgggca 3001 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt 3061 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt 3121 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt 3181 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca 3241 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct 3301 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg 3361 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg 3421 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt 3481 tacagatcca agctgtgacc ggcgcctacg ctagacgcca ccatggagag cgacgagagc 3541 ggcctgatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa 3601 gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga 3661 acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa 3721 atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc 3781 gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc 3841 ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg 3901 ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac 3961 gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa aacggattac 4021 cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa 4081 tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc 4141 tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga 4201 ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta 4261 agtgttgttc cattccatca cggttttgga atgtttacta cactcggata tttgatatgt 4321 ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag 4381 gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc 4441 actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc 4501 ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa 4561 ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa 4621 ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc 4681 gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg 4741 tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta 4801 cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg 4861 aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg 4921 ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt 4981 gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc 5041 gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt 5101 gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc 5161 ctcataaagg ccaagaaggg cggaaagatc gccgtgagat cccgcgctca gtcgtccaat 5221 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt 5281 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg 5341 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 5401 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 5461 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 5521 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 5581 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 5641 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 5701 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 5761 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 5821 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 5881 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa 5941 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 6001 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 6061 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 6121 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 6181 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 6241 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 6301 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 6361 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 6421 cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt 6481 cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 6541 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 6601 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 6661 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 6721 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 6781 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 6841 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 6901 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 6961 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 7021 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 7081 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 7141 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 7201 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 7261 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 7321 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 7381 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 7441 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7501 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7561 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7621 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7681 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7741 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7801 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7861 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7921 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7981 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 8041 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 8101 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 8161 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 8221 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 8281 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 8341 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 8401 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 8461 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 8521 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 8581 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8641 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8701 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8761 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 8821 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8881 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8941 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 9001 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 9061 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 9121 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 9181 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 9241 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 9301 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 9361 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 9421 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 9481 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 9541 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 9601 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 9661 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 9721 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 9781 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //