LOCUS V046081 9670 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046081 VERSION V046081 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..4366 /label="BICDL2(NM_001369667)" /note="BICDL2(NM_001369667)" /gene="BICDL2" CDS 4373..4438 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4458..4957 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4978..5577 /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 5591..6179 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6386..7019 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7148..7164) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7172..7188 /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(7196..7226) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7241..7262 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7550..8138) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8309..9169) /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(9170..9274) /gene="bla" /label="AmpR promoter" polyA_signal 9322..9456 /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 ccatgagctc tccagatggg cccagcttcc cgtccgggcc 2881 gctctcaggg ggcgcctctc ccagcggcga cgagggcttc ttcccctttg tgctggagcg 2941 gcgggactca ttcctgggag ggggcccagg gcctgaggag cccgaggacc tagccttgca 3001 gctgcagcag aaggagaaag acctgctgtt ggccgcggag ctcggcaaga tgcttctgga 3061 gcgaaatgaa gagctgcggc ggcagctgga gacgctgagc gcccagcact tggagcgtga 3121 ggaacggctc cagcaggaga accatgagct ccggcgaggc ctggcagccc gaggagccga 3181 gtgggaggcc cgggccgtgg agctggaggg ggacgtggag gccctgcggg cccagcttgg 3241 ggagcagcgc tcagagcagc aggacagtgg gcgagaacgg gcacgggccc tcagcgagct 3301 cagcgagcag aacctccggc tcagccagca gctggctcag gcctcccaga ctgagcagga 3361 acttcagagg gaactggacg cccttcgggg acagtgccag gctcaggcac tggctggagc 3421 agagctgagg acgcggctgg agagtctgca gggggaaaac cagatgctgc agagccgccg 3481 gcaggacctg gaggcccaga tccgaggcct gcgtgaggag gtggagaagg gtgagggcag 3541 actgcagacc acccacgagg agttgctgct gctgaggcgg gagcggcggg agcacagcct 3601 ggagctggaa cgcgcacggt cagaggctgg ggaggcgctg agtgcgctgc ggaggctgca 3661 gcggcgcgtc tccgagctgg aggaggagtc acgcctccag gacgccgacg tgtcggctgc 3721 ctcgttgcag tcagaactgg cccacagcct cgacgacggc gaccagggcc agggcgccga 3781 cgcacccgga gacaccccga ccacccggtc cccaaagacc cgaaaggcgt ccagccccca 3841 gccttcaccc ccggaagaga tcttagagcc ccccaagaag cgaacatccc tcagtccagc 3901 ggagatactg gaggagaagg aggtggaagt ggccaagctg caagatgaga tctcgctgca 3961 gcaggcagag ctgcagtccc tgcgggaaga gctgcagagg cagaaggagc tgcgggcgca 4021 ggaagaccct ggggaggccc tgcacagtgc cctctcagac cgggacgagg ccgtgaacaa 4081 ggccctggag ctgtccctgc agctcaaccg cgtctcgctg gagcgagact ccctgtctcg 4141 ggagctgctg cgcgccatcc gccagaaggt ggcgctcacg caggagctgg aggcctggca 4201 ggacgacatg caggtggtga tcgggcagca gctgcgctca cagcgccaga aggagctgag 4261 cgcctccgcg tcgtcgtcga ccccgcgccg tgccgcgccc cgcttctcgc tgcgcctggg 4321 ccccgggccc gccggtggct ttctcagtaa cctcttccga aggaccggat ccgactacaa 4381 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 4441 gtctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc 4501 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc 4561 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc 4621 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg 4681 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg 4741 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc 4801 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc 4861 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc 4921 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg 4981 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 5041 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 5101 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 5161 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 5221 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 5281 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 5341 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 5401 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 5461 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 5521 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg 5581 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 5641 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 5701 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 5761 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 5821 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 5881 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 5941 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc 6001 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 6061 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 6121 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 6181 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca 6241 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt 6301 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 6361 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat 6421 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 6481 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 6541 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 6601 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 6661 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 6721 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 6781 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 6841 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 6901 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 6961 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 7021 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 7081 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt 7141 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 7201 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 7261 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 7321 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 7381 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 7441 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 7501 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 7561 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 7621 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 7681 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 7741 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 7801 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 7861 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 7921 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 7981 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 8041 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 8101 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 8161 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 8221 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 8281 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 8341 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 8401 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 8461 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 8521 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 8581 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 8641 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 8701 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 8761 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 8821 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 8881 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 8941 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 9001 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 9061 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 9121 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 9181 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 9241 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 9301 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc 9361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 9421 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 9481 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 9541 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 9601 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa 9661 acttagtagt //