LOCUS V035482 9592 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035482 VERSION V035482 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 2843..4288 /label="MKRN1(NM_013446)" /note="MKRN1(NM_013446)" /gene="MKRN1" CDS 4295..4360 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4380..4879 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4900..5499 /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 5513..6101 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6308..6941 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7070..7086) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7094..7110 /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(7118..7148) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7163..7184 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7472..8060) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8231..9091) /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(9092..9196) /gene="bla" /label="AmpR promoter" polyA_signal 9244..9378 /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 ctcaagcttc gaattcgcca ccatggcgga ggctgcaact cccggaacaa cagccacaac 2881 atcaggagca ggagcggcag cggcgacggc ggcagcagcc tcccccaccc cgatccccac 2941 agtcaccgcc ccgtccctgg gggcgggcgg agggggcggc ggcagcgacg gcagcggcgg 3001 cggctggact aaacaggtca cctgcaggta ttttatgcat ggggtttgta aggaaggaga 3061 caactgtcgc tactcgcatg acctctctga cagtccgtat agtgtagtgt gcaagtattt 3121 tcagcgaggg tactgtattt atggagaccg ctgcagatat gaacatagca aaccattgaa 3181 acaggaagaa gcaactgcta cagagctaac tacaaagtca tcccttgctg cttcctcaag 3241 tctctcatcg atagttggac cacttgttga aatgaataca ggcgaagctg agtcaagaaa 3301 ttcaaacttt gcaactgtag gagcaggttc agaggactgg gtgaatgcta ttgagtttgt 3361 tcctgggcaa ccctactgtg gccgtactgc gccttcctgc actgaagcac ccctgcaggg 3421 ctcagtgacc aaggaagaat cagagaaaga gcaaaccgcc gtggagacaa agaagcagct 3481 gtgcccctat gctgcagtgg gagagtgccg atacggggag aactgtgtgt atctccacgg 3541 agattcttgt gacatgtgtg ggctgcaggt cctgcatcca atggatgctg cccagagatc 3601 gcagcatatc aaatcgtgca ttgaggccca tgagaaggac atggagctct catttgccgt 3661 gcagcgcagc aaggacatgg tgtgtgggat ctgcatggag gtggtctatg agaaagccaa 3721 ccccagtgag cgccgcttcg ggatcctctc caactgcaac cacacctact gtctcaagtg 3781 cattcgcaag tggaggagtg ctaagcaatt tgagagcaag atcataaagt cctgcccaga 3841 atgccggatc acatctaact ttgtcattcc aagtgagtac tgggtggagg agaaagaaga 3901 gaagcagaaa ctcattctga aatacaagga ggcaatgagc aacaaggcgt gcaggtattt 3961 tgatgaagga cgtgggagct gcccatttgg agggaactgt ttttacaagc atgcgtaccc 4021 tgatggccgt agagaggagc cacagagaca gaaagtggga acatcaagca gataccgggc 4081 ccaacgaagg aaccacttct gggaactcat tgaggaaaga gagaacagca acccctttga 4141 caacgatgaa gaagaggttg tcacctttga gctgggcgag atgttgctta tgcttttggc 4201 tgcaggtggg gacgacgaac taacagactc tgaagatgag tgggacttgt ttcatgatga 4261 gctggaagat ttttatgact tggatctagg atccgactac aaagaccatg acggtgatta 4321 taaagatcat gacatcgact acaaggatga cgatgacaag tagtctagat aattctaccg 4381 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg 4441 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg 4501 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag 4561 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg 4621 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg 4681 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg 4741 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc 4801 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc 4861 ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg 4921 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 4981 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 5041 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 5101 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 5161 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 5221 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 5281 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 5341 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 5401 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 5461 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac cgcgtctgga acaatcaacc 5521 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 5581 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 5641 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 5701 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 5761 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 5821 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 5881 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt 5941 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 6001 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 6061 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt 6121 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat 6181 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta 6241 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 6301 aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc 6361 taccacacac aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga 6421 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa 6481 gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat 6541 gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg 6601 gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga 6661 ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc 6721 ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca 6781 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6841 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6901 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6961 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca 7021 ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 7081 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 7141 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 7201 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 7261 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 7321 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 7381 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 7441 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 7501 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 7561 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 7621 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 7681 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 7741 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 7801 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 7861 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 7921 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 7981 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 8041 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 8101 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 8161 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 8221 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 8281 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 8341 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 8401 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 8461 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 8521 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 8581 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 8641 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 8701 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 8761 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 8821 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 8881 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 8941 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 9001 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 9061 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 9121 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 9181 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 9241 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 9301 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 9361 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc 9421 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat 9481 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta 9541 gtagttttta aagaaattgt atttgttaaa tatgtactac aaacttagta gt //