LOCUS V027936 9466 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027936 VERSION V027936 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 2826..4169 /label="Azin1(NM_001102458)" /note="Azin1(NM_001102458)" /gene="Azin1" CDS 4170..4193 /codon_start=1 /product="8xHis affinity tag" /transl_table=1 /translation="HHHHHHHH" /label="8xHis" promoter 4254..4753 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4774..5373 /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 5387..5975 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6182..6815 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6944..6960) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6968..6984 /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(6992..7022) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7037..7058 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7346..7934) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8105..8965) /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(8966..9070) /gene="bla" /label="AmpR promoter" polyA_signal 9118..9252 /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 tcgctagcgc taccggactc agatctcgag 2821 ccaccatgaa aggatttatt gacgatgcga actactccgt tggcctgttg gatgaaggaa 2881 caaaccttgg aaatgttatt gataactatg tttatgaaca taccctgaca ggaaaaaatg 2941 cattttttgt gggggatctt gggaagatcg tgaagaagca cagtcagtgg cagaccgtgg 3001 tggctcagat aaagccgttt tacacggtga agtgcaactc cactccagcc gtgcttgaga 3061 tcttggcagc tcttggaact gggtttgctt gttccagcaa aaatgaaatg gctttagtgc 3121 aagaattggg tgtatctcca gaaaacatca ttttcacaag tccttgtaag caagtgtctc 3181 agataaagta tgcagcaaaa gttggagtaa atattatgac atgtgacaat gagattgaat 3241 taaagaaaat tgcaaggaat cacccaaatg ccaaggtctt actacatatt gcaacagaag 3301 ataatattgg aggtgaagat ggtaacatga agtttggcac tacactgaag aattgtaggc 3361 atcttttgga atgtgccaag gaacttgatg tccaaataat tggggttaaa tttcatgttt 3421 caagtgcttg caaagaatat caagtatatg tacatgccct gtctgatgct cgatgtgtgt 3481 ttgacatggc tggagagttt ggctttacaa tgaacatgtt agacatcggt ggaggcttca 3541 caggaactga aattcagttg gaagaggtta atcatgttat cagtcctctg ttggatattt 3601 acttccctga aggatctggc attcagataa tttcagaacc tggaagctac tatgtatctt 3661 ctgcgtttac acttgcagtc aatattattg ctaagaaagt tgttgaaaat gataaatttt 3721 cctctggagt agaaaaaaat gggagtgatg agccagcctt cgtgtattac atgaatgatg 3781 gtgtttatgg ttcttttgcg agtaagcttt ctgaggactt aaataccatt ccagaggttc 3841 acaagaaata caaggaagat gagcctctgt ttacaagcag cctttggggt ccatcctgtg 3901 atgagcttga tcaaattgtg gaaagctgtc ttcttcctga gctgaatgtg ggagattggc 3961 ttatctttga taacatggga gcagattctt tccacgaacc atctgctttt aatgattttc 4021 agaggccagc tatttatttc atgatgtcat tcagtgattg gtatgagatg caagatgctg 4081 gaattacttc agatgcaatg atgaaaaact tcttctttgc accctcttgt attcagctga 4141 gccaagaaga cagcttttcc actgaagctc accatcacca tcaccatcac cattgaattc 4201 tgcagtcgac ggtaccgcgg gcccgggatc ccgcgactct agataattct accgggtagg 4261 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg 4321 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc 4381 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt 4441 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac 4501 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag 4561 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc 4621 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg 4681 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc 4741 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc 4801 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac 4861 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg 4921 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac 4981 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc 5041 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg 5101 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc 5161 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag 5221 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc 5281 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc 5341 acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga 5401 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 5461 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 5521 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 5581 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 5641 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 5701 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 5761 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac 5821 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 5881 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 5941 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac 6001 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc 6061 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta 6121 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga 6181 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac 6241 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca 6301 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc 6361 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg 6421 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga 6481 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc 6541 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga 6601 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg 6661 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 6721 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 6781 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat 6841 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta 6901 gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 6961 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 7021 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 7081 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 7141 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 7201 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 7261 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 7321 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 7381 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 7441 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 7501 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 7561 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 7621 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 7681 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 7741 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 7801 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 7861 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 7921 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 7981 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 8041 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 8101 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 8161 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 8221 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 8281 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 8341 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 8401 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 8461 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 8521 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 8581 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 8641 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 8701 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 8761 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 8821 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 8881 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 8941 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 9001 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 9061 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac 9121 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 9181 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 9241 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca 9301 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct 9361 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt 9421 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt //