LOCUS V039738 9860 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039738 VERSION V039738 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" regulatory 2820..2829 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2829..2894 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2901..4616 /label="DPYSL2(NM_001386)" /note="DPYSL2(NM_001386)" /gene="DPYSL2" promoter 4648..5147 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5168..5767 /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 5781..6369 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6576..7209 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7338..7354) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7362..7378 /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(7386..7416) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7431..7452 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7740..8328) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8499..9359) /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(9360..9464) /gene="bla" /label="AmpR promoter" polyA_signal 9512..9646 /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 ccaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg 2881 atgacgatga caaggaattc atgtcttatc aggggaagaa aaatattcca cgcatcacga 2941 gcgatcgtct tctgatcaaa ggaggtaaaa ttgttaatga tgaccagtcg ttctatgcag 3001 acatatacat ggaagatggg ttgatcaagc aaataggaga aaatctgatt gtgccaggag 3061 gagtgaagac catcgaggcc cactcccgga tggtgatccc cggaggaatt gacgtccaca 3121 ctcgtttcca gatgcctgat cagggaatga cgtctgctga tgatttcttc caaggaacca 3181 aggcggccct ggctggggga accactatga tcattgacca cgttgttcct gagcctggga 3241 caagcctgct cgctgccttt gaccagtgga gggaatgggc cgacagcaag tcctgctgtg 3301 actactctct gcatgtggac atcagtgagt ggcataaggg catccaggag gagatggaag 3361 cgcttgtgaa ggatcacggg gtaaattcct tcctcgtgta catggctttc aaagatcgct 3421 tccagctaac ggattgccag atttatgaag tactgagtgt gatccgggat attggcgcca 3481 tagcccaagt ccacgcagaa aatggcgaca tcattgcaga ggagcagcag aggatcctgg 3541 atctgggcat cacgggcccc gagggacatg tgctgagccg acctgaggag gtcgaggccg 3601 aagccgtgaa tcgtgccatc accatcgcca accagaccaa ctgcccgctg tatatcacca 3661 aggtgatgag caaaagctct gctgaggtca tcgcccaggc acggaagaag ggaactgtgg 3721 tgtatggcga gcccatcact gccagcttgg gaacggacgg ctcccattac tggagcaaga 3781 actgggccaa ggctgctgcc tttgtcacct ccccaccctt gagccctgat ccaaccactc 3841 cagactttct caactccttg ctgtcctgtg gagacctcca ggtcacgggc agtgcccatt 3901 gcacgtttaa cactgcccag aaggctgtag gaaaggacaa cttcaccctg attccggagg 3961 gcaccaatgg cactgaggag cggatgtccg tcatctggga caaggctgtg gtcactggga 4021 agatggatga gaaccagttt gtggctgtga ccagcaccaa tgcagccaaa gtcttcaacc 4081 tttacccccg gaaaggccgc attgctgtgg gatccgatgc cgacctggtc atctgggacc 4141 ccgacagcgt taaaaccatc tctgccaaga cacacaacag ctctctcgag tacaacatct 4201 ttgaaggcat ggagtgccgc ggctccccac tggtggtcat cagccagggg aagattgtcc 4261 tggaggacgg caccctgcat gtcaccgaag gctctggacg ctacattccc cggaagccct 4321 tccctgattt tgtttacaag cgtatcaagg caaggagcag gctggctgag ctgagagggg 4381 ttcctcgtgg cctgtatgac ggacctgtgt gtgaagtgtc tgtgacgccc aagacagtca 4441 ctccagcctc ctcggccaag acgtctcctg ccaagcagca ggccccacct gtccggaacc 4501 tgcaccagtc tggattcagt ttgtctggtg ctcagattga tgacaacatt ccccgccgca 4561 ccacccagcg tatcgtggcg ccccccggtg gccgtgccaa catcaccagc ctgggctagg 4621 gatcccgcga ctctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg 4681 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc 4741 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg 4801 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt 4861 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg 4921 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg 4981 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca 5041 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc 5101 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa 5161 gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag 5221 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga 5281 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg 5341 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac 5401 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt 5461 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc 5521 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg 5581 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc 5641 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt 5701 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg 5761 tgcctgaccg cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg 5821 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt 5881 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc 5941 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt 6001 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga 6061 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct 6121 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga 6181 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct 6241 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc 6301 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg 6361 cctccccgcc tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag 6421 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga 6481 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt 6541 agatcttagc cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg 6601 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca 6661 gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct 6721 agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt 6781 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt 6841 tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa 6901 ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg 6961 cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg 7021 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag 7081 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc 7141 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa 7201 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat 7261 gaatatcaga gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag 7321 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 7381 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 7441 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 7501 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 7561 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 7621 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 7681 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 7741 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 7801 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 7861 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 7921 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 7981 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 8041 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 8101 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 8161 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 8221 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 8281 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 8341 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 8401 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 8461 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 8521 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 8581 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 8641 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 8701 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 8761 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 8821 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 8881 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 8941 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 9001 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 9061 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 9121 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 9181 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 9241 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 9301 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 9361 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 9421 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 9481 gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta 9541 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 9601 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca 9661 agctaaccaa aatcatccca aacttcccac cccataccct attaccactg ccaattacct 9721 gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg 9781 tatttgttaa atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata 9841 tgtactacaa acttagtagt //