LOCUS V021225 9106 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021225 VERSION V021225 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 2829..2855 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2856..3863 /label="XRCC4(NM_022406)" /note="XRCC4(NM_022406)" /gene="XRCC4" promoter 3894..4393 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4414..5013 /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 5027..5615 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5822..6455 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6584..6600) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6608..6624 /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(6632..6662) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6677..6698 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6986..7574) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7745..8605) /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(8606..8710) /gene="bla" /label="AmpR promoter" polyA_signal 8758..8892 /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 ccaccatgta cccatacgac gtaccagatt acgctatgga gagaaaaata agcagaatcc 2881 accttgtttc tgaacccagt ataactcatt ttctacaagt atcttgggag aaaacactgg 2941 aatctggttt tgttattaca cttactgatg gtcattcagc atggactggg acagtttctg 3001 aatcagagat ttcccaagaa gctgatgaca tggcaatgga aaaagggaaa tatgttggtg 3061 aactgagaaa agcattgttg tcaggagcag gaccagctga tgtatacacg tttaattttt 3121 ctaaagagtc ttgttatttc ttctttgaga aaaacctgaa agatgtctca ttcagacttg 3181 gttccttcaa cctagagaaa gttgaaaacc cagctgaagt cattagagaa cttatttgtt 3241 attgcttgga caccattgca gaaaatcaag ccaaaaatga gcacctgcag aaagaaaatg 3301 aaaggcttct gagagattgg aatgatgttc aaggacgatt tgaaaaatgt gtgagtgcta 3361 aggaagcttt ggagactgat ctttataagc ggtttattct ggtgttgaat gagaagaaaa 3421 caaaaatcag aagtttgcat aataaattat taaatgcagc tcaagaacga gaaaaggaca 3481 tcaaacaaga aggggaaact gcaatctgtt ctgaaatgac tgctgaccga gatccagtct 3541 atgatgagag tactgatgag gaaagtgaaa accaaactga tctctctggg ttggcttcag 3601 ctgctgtaag taaagatgat tccattattt caagtcttga tgtcactgat attgcaccaa 3661 gtagaaaaag gagacagcga atgcaaagaa atcttgggac agaacctaaa atggctcctc 3721 aggagaatca gcttcaagaa aaggaaaatt ctaggcctga ttcttcacta cctgagacgt 3781 ctaaaaagga gcacatctca gctgaaaaca tgtctttaga aactctgaga aacagcagcc 3841 cagaagacct ctttgatgag atttaggatc ccgcgactct agataattct accgggtagg 3901 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg 3961 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc 4021 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt 4081 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac 4141 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag 4201 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc 4261 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg 4321 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc 4381 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc 4441 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac 4501 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg 4561 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac 4621 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc 4681 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg 4741 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc 4801 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag 4861 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc 4921 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc 4981 acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga 5041 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 5101 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 5161 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 5221 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 5281 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 5341 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 5401 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac 5461 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 5521 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 5581 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac 5641 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc 5701 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta 5761 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga 5821 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac 5881 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca 5941 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc 6001 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg 6061 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga 6121 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc 6181 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga 6241 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg 6301 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 6361 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 6421 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat 6481 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta 6541 gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 6601 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 6661 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 6721 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 6781 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 6841 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 6901 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 6961 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 7021 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 7081 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 7141 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 7201 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 7261 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 7321 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 7381 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 7441 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 7501 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 7561 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 7621 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 7681 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 7741 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 7801 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 7861 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 7921 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 7981 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 8041 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 8101 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 8161 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 8221 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 8281 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 8341 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 8401 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 8461 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 8521 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 8581 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 8641 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 8701 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac 8761 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 8821 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 8881 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca 8941 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct 9001 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt 9061 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt //