LOCUS V019770 8522 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019770 VERSION V019770 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..3278 /label="Lect2(NM_010702)" /note="Lect2(NM_010702)" /gene="Lect2" promoter 3310..3809 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 3830..4429 /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 4443..5031 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5238..5871 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6000..6016) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6024..6040 /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(6048..6078) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6093..6114 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6402..6990) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7161..8021) /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(8022..8126) /gene="bla" /label="AmpR promoter" polyA_signal 8174..8308 /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 ccaccatgat tcccacaaca atcctcattt cagctgcttt gctttcctct gccctagcag 2881 gaccatgggc taacatatgt gccagcaaat cttccaacga gatccggacg tgtgacagct 2941 atggctgtgg acagtactct gctcaaagaa cccaaaggca tcacccaggt gtggacgtcc 3001 tgtgctcgga tggatctgtg gtgtatgcac cattcactgg gaagatagtg ggccaggaga 3061 aaccctatag aaacaaaaat gccatcaatg atggcattcg actgtctgga agaggttttt 3121 gtgtcaaaat tttctacatt aagccaatta agtataaagg ttctatcaaa aagggggaga 3181 agctgggcac cttgctgccc ctgcagaaag tttacccggg catccagtcg catgtacacg 3241 ttgaaaactg cgactccagt gaccccacag catacctgta aggatcccgc gactctagat 3301 aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag 3361 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca 3421 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc 3481 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg 3541 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg 3601 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc 3661 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc 3721 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt 3781 tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta 3841 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc 3901 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga 3961 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt 4021 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc 4081 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc 4141 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt 4201 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt 4261 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc 4321 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt 4381 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac cgcgtctgga 4441 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 4501 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 4561 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 4621 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 4681 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 4741 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 4801 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc 4861 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 4921 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 4981 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta 5041 attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac 5101 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac 5161 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt 5221 aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga 5281 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc 5341 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga 5401 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca 5461 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt 5521 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg 5581 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga 5641 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct 5701 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc 5761 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg 5821 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt 5881 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga 5941 ggccttgaca ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg 6001 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 6061 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 6121 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 6181 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 6241 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 6301 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 6361 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 6421 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 6481 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 6541 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 6601 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 6661 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 6721 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 6781 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 6841 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 6901 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 6961 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 7021 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 7081 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 7141 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 7201 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 7261 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 7321 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 7381 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 7441 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 7501 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 7561 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 7621 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 7681 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 7741 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 7801 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 7861 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 7921 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 7981 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 8041 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 8101 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac 8161 ggatcgggag atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 8221 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 8281 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc 8341 caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta 8401 aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac 8461 tacaaactta gtagttttta aagaaattgt atttgttaaa tatgtactac aaacttagta 8521 gt //