LOCUS V039217 10402 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039217 VERSION V039217 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..5089 /label="LIN54(NM_194282)" /note="LIN54(NM_194282)" /gene="LIN54" CDS 5105..5170 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 5190..5689 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5710..6309 /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 6323..6911 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7118..7751 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7880..7896) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7904..7920 /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(7928..7958) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7973..7994 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8282..8870) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9041..9901) /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(9902..10006) /gene="bla" /label="AmpR promoter" polyA_signal 10054..10188 /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 ccatggaggt ggtgccagct gaggtgaata gtttgcttcc 2881 agaggaaata atggacactg gtataacttt agtggatgat gatagtattg aggctgttat 2941 tgtttcatcc ccaattccca tggagacaga actggaagaa attgtcaaca taaattctac 3001 tggtgactct acagccacgc ccatttccac ggaaccaatc acagtgtaca gtaaccacac 3061 taaccaagtt gcagtgaata ccacaattac taaagcagat tctaatacca cagtgaaacc 3121 agcttttcca agtggccttc aaaaacttgg tgctcagact cctgtgacta tatcagccaa 3181 tcagattatt ttaaacaaag tatcacagac atctgatctt aaacttggca atcagaccct 3241 taaaccagat ggacagaagt taattttaac aactttgggc aagtctggtt caccaattgt 3301 tttagcacta ccccatagcc aactacccca ggctcagaaa gttacaactc aggcccagtc 3361 aggagatgct aagttaccac cgcagcaaat taaagtagtt accattggag ggaggccaga 3421 ggtgaaacct gtcattggtg tctcagcatt gaccccagga agtcaactga ttaatactac 3481 aactcagccc tctgtgttac agacccaaca gttaaaaaca gtacagattg ctaagaagcc 3541 tcgaacgcca acctctggtc cagtaatcac gaagctgatc tttgcaaaac caattaatag 3601 taaagcagtt acaggacaga caactcaagt ttcaccacca gttattgcag gtagggttct 3661 ttcacagtct actcccggaa ctccatcaaa gaccataaca atatctgaaa gtggtgttat 3721 tggatcaact ttaaattcta caacacagac accaaataaa atagccatct cacctttgaa 3781 atcgccaaat aaggcagtga aatcaactgt gcagaccatc actgttggag gagtgagcac 3841 atcacagttt aagacaatta ttcctctggc aactgctccc aatgtccagc agattcaagt 3901 gcctggaagc aagtttcatt atgtccgact tgttactgcc acatcagcca gtagctcaac 3961 ccagccagtt agtcagaatc ccagtacaaa cactcagcct cttcagcaag caaagccagt 4021 ggttgttaat acaaccccag tgcggatgtc agttccaatt gtctcagctc aggctgtcaa 4081 acaagttgtt ccaaaaccaa tcaatccaac ttcacaaata gtaactacta gccagccaca 4141 gcaacggctt atcatgcctg ccacaccact gccacagatc cagcccaacc tcactaacct 4201 gccaccaggc actgtcctgg caccagctcc gggaacaggg aatgtgggtt atgcagtgct 4261 tccagctcag tatgttactc agctacagca gtcttcatat gtatcaatag caagcaactc 4321 tacctttact ggaacatctg gtatccagac ccaggcacgg cttccattca atggcataat 4381 cccatcagag tcggccagtc ggccccgaaa gccctgtaat tgtacaaaat cactgtgttt 4441 gaaattgtat tgtgattgct ttgcaaatgg tgaattttgc aacaactgca attgtactaa 4501 ttgttacaac aatttggaac atgaaaatga aaggcaaaaa gcaataaagg catgccttga 4561 cagaaatcca gaagccttta agcctaagat agggaaagga aaggagggag aatctgatcg 4621 acgtcatagc aaagggtgta attgcaaacg atcaggatgt cttaaaaact actgtgaatg 4681 ctatgaggca aaaataatgt gttcctcaat atgcaaatgt attggctgta agaattttga 4741 agaaagcccg gaaaggaaga cattgatgca tttggcagat gcagctgaag taagggtaca 4801 gcaacaaaca gcagccaaga cgaagttatc ctctcaaatt tcagacttgc ttactaggcc 4861 aacaccagct ttaaatagtg gaggcggaaa attgccattt acatttgtaa ctaaggaagt 4921 agctgaagcc acatgtaatt gcctccttgc ccaggcagag caggcagaca agaagggaaa 4981 atcaaaggca gcagcggaac ggatgatact tgaggaattc ggacgatgtt tgatgagtgt 5041 catcaactct gcaggaaagg caaaaagtga cccttgtgcc atgaattgcg gtggaggcgg 5101 atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga 5161 cgatgacaag tagtctagat aattctaccg ggtaggggag gcgcttttcc caaggcagtc 5221 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc 5281 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt 5341 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc 5401 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac 5461 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt 5521 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct 5581 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa 5641 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc 5701 aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 5761 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 5821 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 5881 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 5941 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 6001 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 6061 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 6121 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 6181 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 6241 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 6301 ggtgcctgac cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac 6361 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 6421 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 6481 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 6541 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 6601 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 6661 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct 6721 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 6781 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 6841 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 6901 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca 6961 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag 7021 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct 7081 gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa 7141 cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattag 7201 cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag 7261 ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg 7321 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg 7381 tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag 7441 aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt 7501 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt 7561 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 7621 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 7681 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 7741 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa 7801 atgaatatca gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg acctctagct 7861 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7921 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7981 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 8041 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 8101 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 8161 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 8221 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 8281 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 8341 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 8401 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 8461 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 8521 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 8581 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 8641 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 8701 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 8761 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 8821 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8881 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8941 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 9001 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 9061 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 9121 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 9181 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 9241 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 9301 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 9361 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 9421 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 9481 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 9541 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 9601 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 9661 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 9721 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 9781 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9841 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9901 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9961 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 10021 aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc ttataatggt 10081 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 10141 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact 10201 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac 10261 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat 10321 tgtatttgtt aaatatgtac tacaaactta gtagttttta aagaaattgt atttgttaaa 10381 tatgtactac aaacttagta gt //