LOCUS V022466 8679 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022466 VERSION V022466 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 2827..3405 /label="BCL2L2(NM_004050)" /note="BCL2L2(NM_004050)" /gene="BCL2L2" CDS 3412..3435 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 3467..3966 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 3987..4586 /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 4600..5188 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5395..6028 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6157..6173) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6181..6197 /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(6205..6235) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6250..6271 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6559..7147) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7318..8178) /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(8179..8283) /gene="bla" /label="AmpR promoter" polyA_signal 8331..8465 /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 gccaccatgg cgaccccagc ctcggcccca gacacacggg ctctggtggc agactttgta 2881 ggttataagc tgaggcagaa gggttatgtc tgtggagctg gccccgggga gggcccagca 2941 gctgacccgc tgcaccaagc catgcgggca gctggagatg agttcgagac ccgcttccgg 3001 cgcaccttct ctgatctggc ggctcagctg catgtgaccc caggctcagc ccaacaacgc 3061 ttcacccagg tctccgatga actttttcaa gggggcccca actggggccg ccttgtagcc 3121 ttctttgtct ttggggctgc actgtgtgct gagagtgtca acaaggagat ggaaccactg 3181 gtgggacaag tgcaggagtg gatggtggcc tacctggaga cgcagctggc tgactggatc 3241 cacagcagtg ggggctgggc ggagttcaca gctctatacg gggacggggc cctggaggag 3301 gcgcggcgtc tgcgggaggg gaactgggca tcagtgagga cagtgctgac gggggccgtg 3361 gcactggggg ccctggtaac tgtaggggcc ttttttgcta gcaaggaatt cgattacaag 3421 gatgacgacg ataagtgagg atcccgcgac tctagataat tctaccgggt aggggaggcg 3481 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac 3541 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg 3601 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc 3661 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc 3721 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa 3781 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc 3841 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca 3901 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc 3961 ctttcgacct gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca 4021 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg 4081 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac 4141 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg 4201 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg 4261 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc 4321 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc 4381 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg 4441 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg 4501 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt 4561 gcatgacccg caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa 4621 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 4681 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 4741 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 4801 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 4861 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 4921 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 4981 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 5041 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 5101 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 5161 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa 5221 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag 5281 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa 5341 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag 5401 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag 5461 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct 5521 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag 5581 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag 5641 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc 5701 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg 5761 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca 5821 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg 5881 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 5941 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 6001 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat 6061 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgtttt 6121 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 6181 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 6241 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 6301 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 6361 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 6421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 6481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 6541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 6601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 6661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 6721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 6781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 6841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 6901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 6961 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 7021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 7081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 7141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 7201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 7261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 7321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 7381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 7441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 7501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 7561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 7621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 7681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 7741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 7801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 7861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 7921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 7981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 8041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 8101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 8161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 8221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 8281 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta 8341 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 8401 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 8461 ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta 8521 ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat 8581 tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta gtttttaaag 8641 aaattgtatt tgttaaatat gtactacaaa cttagtagt //