LOCUS V027743 9727 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027743 VERSION V027743 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..4423 /label="RAG2(NM_000536)" /note="RAG2(NM_000536)" /gene="RAG2" CDS 4430..4495 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4515..5014 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5035..5634 /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 5648..6236 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6443..7076 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7205..7221) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7229..7245 /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(7253..7283) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7298..7319 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7607..8195) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8366..9226) /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(9227..9331) /gene="bla" /label="AmpR promoter" polyA_signal 9379..9513 /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 ccatgtctct gcagatggta acagtcagta ataacatagc 2881 cttaattcag ccaggcttct cactgatgaa ttttgatgga caagttttct tctttggaca 2941 aaaaggctgg cccaaaagat cctgccccac tggagttttc catctggatg taaagcataa 3001 ccatgtcaaa ctgaagccta caattttctc taaggattcc tgctacctcc ctcctcttcg 3061 ctacccagcc acttgcacat tcaaaggcag cttggagtct gaaaagcatc aatacatcat 3121 ccatggaggg aaaacaccaa acaatgaggt ttcagataag atttatgtca tgtctattgt 3181 ttgcaagaac aacaaaaagg ttacttttcg ctgcacagag aaagacttgg taggagatgt 3241 tcctgaagcc agatatggtc attccattaa tgtggtgtac agccgaggga aaagtatggg 3301 tgttctcttt ggaggacgct catacatgcc ttctacccac agaaccacag aaaaatggaa 3361 tagtgtagct gactgcctgc cctgtgtttt cctggtggat tttgaatttg ggtgtgctac 3421 atcatacatt cttccagaac ttcaggatgg gctatctttt catgtctcta ttgccaaaaa 3481 tgacaccatc tatattttag gaggacattc acttgccaat aatatccggc ctgccaacct 3541 gtacagaata agggttgatc ttcccctggg tagcccagct gtgaattgca cagtcttgcc 3601 aggaggaatc tctgtctcca gtgcaatcct gactcaaact aacaatgatg aatttgttat 3661 tgttggtggc tatcagcttg aaaatcaaaa aagaatgatc tgcaacatca tctctttaga 3721 ggacaacaag atagaaattc gtgagatgga gaccccagat tggaccccag acattaagca 3781 cagcaagata tggtttggaa gcaacatggg aaatggaact gtttttcttg gcataccagg 3841 agacaataaa caagttgttt cagaaggatt ctatttctat atgttgaaat gtgctgaaga 3901 tgatactaat gaagagcaga caacattcac aaacagtcaa acatcaacag aagatccagg 3961 ggattccact ccctttgaag actctgaaga attttgtttc agtgcagaag caaatagttt 4021 tgatggtgat gatgaatttg acacctataa tgaagatgat gaagaagatg agtctgagac 4081 aggctactgg attacatgct gccctacttg tgatgtggat atcaacactt gggtaccatt 4141 ctattcaact gagctcaaca aacccgccat gatctactgc tctcatgggg atgggcactg 4201 ggtccatgct cagtgcatgg atctggcaga acgcacactc atccatctgt cagcaggaag 4261 caacaagtat tactgcaatg agcatgtgga gatagcaaga gctctacaca ctccccaaag 4321 agtcctaccc ttaaaaaagc ctccaatgaa atccctccgt aaaaaaggtt ctggaaaaat 4381 cttgactcct gccaagaaat cctttcttag aaggttgttt gatggatccg actacaaaga 4441 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtc 4501 tagataattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt 4561 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac 4621 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac 4681 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca 4741 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa 4801 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca 4861 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg 4921 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc 4981 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc 5041 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 5101 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 5161 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 5221 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 5281 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5341 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5401 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5461 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5521 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5581 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaccgcgt 5641 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5701 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5761 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5821 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 5881 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 5941 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 6001 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg 6061 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 6121 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 6181 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 6241 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca 6301 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca 6361 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 6421 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc 6481 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca 6541 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag 6601 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc 6661 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta 6721 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga 6781 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact 6841 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc 6901 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 6961 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 7021 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 7081 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 7141 tgagaggcct tgacattgct agcgttttac cgtcgacctc tagctagagc ttggcgtaat 7201 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 7261 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 7321 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 7381 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 7441 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 7501 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 7561 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 7621 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 7681 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 7741 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 7801 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 7861 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 7921 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 7981 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 8041 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 8101 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 8161 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 8221 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 8281 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 8341 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 8401 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 8461 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 8521 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 8581 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 8641 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 8701 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 8761 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 8821 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 8881 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 8941 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 9001 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 9061 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 9121 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 9181 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 9241 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 9301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 9361 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat 9421 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 9481 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat 9541 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta 9601 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata 9661 tgtactacaa acttagtagt ttttaaagaa attgtatttg ttaaatatgt actacaaact 9721 tagtagt //