LOCUS V028548 9655 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028548 VERSION V028548 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2815..4302 /label="NOTUM(NM_178493)" /note="NOTUM(NM_178493)" /gene="NOTUM" CDS 4303..4320 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" misc_feature 4358..4944 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 4964..5563 /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 5577..6165 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6372..7005 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7133..7149) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7157..7173 /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(7181..7211) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7226..7247 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7535..8123) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8294..9154) /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(9155..9259) /gene="bla" /label="AmpR promoter" polyA_signal 9307..9441 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatgggc 2821 cgaggggtgc gcgtgctgct gctgctgagc ctgctgcact gcgccggggg cagcgagggc 2881 aggaagacct ggcggcgccg gggtcagcag ccgcctcctc ccccgcggac cgaggcggcg 2941 ccggcggccg gacagcccgt ggagagcttc ccgctggact tcacggccgt ggagggtaac 3001 atggacagct tcatggcgca agtcaagagc ctggcgcagt ccctgtaccc ctgctccgcg 3061 cagcagctca acgaggacct gcgcctgcac ctcctactca acacctcggt gacctgcaac 3121 gacggcagcc ccgccggcta ctacctgaag gagtccaggg gcagccggcg gtggctcctc 3181 ttcctggaag gcggctggta ctgcttcaac cgcgagaact gcgactccag atacgacacc 3241 atgcggcgcc tcatgagctc ccgggactgg ccgcgcactc gcacaggcac agggatcctg 3301 tcctcacagc cggaggagaa cccctactgg tggaacgcaa acatggtctt catcccctac 3361 tgctccagtg atgtttggag cggggcttca tccaagtctg agaagaacga gtacgccttc 3421 atgggcgccc tcatcatcca ggaggtggtg cgggagcttc tgggcagagg gctgagcggg 3481 gccaaggtgc tgctgctggc cgggagcagc gcggggggca ccggggtgct cctgaatgtg 3541 gaccgtgtgg ctgagcagct ggagaagctg ggctacccag ccatccaggt gcgaggcctg 3601 gctgactccg gctggttcct ggacaacaag cagtatcgcc acacagactg cgtcgacacg 3661 atcacgtgcg cgcccacgga ggccatccgc cgtggcatca ggtactggaa cggggtggtc 3721 ccggagcgct gccgacgcca gttccaggag ggcgaggagt ggaactgctt ctttggctac 3781 aaggtctacc cgaccctgcg ctgccctgtg ttcgtggtgc agtggctgtt tgacgaggca 3841 cagctgacgg tggacaacgt gcacctgacg gggcagccgg tgcaggaggg cctgcggctg 3901 tacatccaga acctcggccg cgagctgcgc cacacactca aggacgtgcc ggccagcttt 3961 gcccccgcct gcctctccca tgagatcatc atccggagcc actggacgga tgtccaggtg 4021 aaggggacgt cgctgccccg agcactgcac tgctgggaca ggagcctcca tgacagccac 4081 aaggccagca agacccccct caagggctgc cccgtccacc tggtggacag ctgcccctgg 4141 ccccactgca acccctcatg ccccaccgtc cgagaccagt tcacggggca agagatgaac 4201 gtggcccagt tcctcatgca catgggcttc gacatgcaga cggtggccca gccgcaggga 4261 ctggagccca gtgagctgct ggggatgctg agcaacggaa gccatcatca ccatcaccat 4321 tagctcgaga ctagttctag agcggccgcg gatcccgccc ctctccctcc ccccccccta 4381 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt 4441 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga 4501 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg 4561 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt 4621 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat 4681 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg 4741 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg 4801 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt 4861 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa 4921 cacgatgata agcttgccac aacccacaag gagacgacct tccatgaccg agtacaagcc 4981 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc 5041 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt 5101 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 5161 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 5221 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 5281 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 5341 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 5401 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 5461 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga 5521 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tagacgcgtc tggaacaatc 5581 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 5641 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 5701 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 5761 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 5821 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 5881 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 5941 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 6001 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 6061 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 6121 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg 6181 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc 6241 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca 6301 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 6361 aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg 6421 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt 6481 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt 6541 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat 6601 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca 6661 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa 6721 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg 6781 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag 6841 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat 6901 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact 6961 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc 7021 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt 7081 gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 7141 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 7201 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 7261 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 7321 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 7381 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 7441 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 7501 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 7561 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 7621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 7681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 7741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 7801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 7861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 7921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 7981 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 8041 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 8101 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 8161 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 8221 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 8281 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 8341 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 8401 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 8461 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 8521 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 8581 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 8641 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 8701 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 8761 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 8821 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 8881 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 8941 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 9001 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 9061 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 9121 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 9181 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 9241 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 9301 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 9361 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 9421 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt 9481 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt 9541 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac 9601 ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //