LOCUS V028182 10870 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028182 VERSION V028182 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..5605 /label="RNF31(NM_001310332)" /note="RNF31(NM_001310332)" /gene="RNF31" CDS 5612..5638 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 5658..6157 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 6178..6777 /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 6791..7379 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7586..8219 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8348..8364) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8372..8388 /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(8396..8426) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8441..8462 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8750..9338) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9509..10369) /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(10370..10474) /gene="bla" /label="AmpR promoter" polyA_signal 10522..10656 /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 ccatggactt atgcaccagg gctggggagc ccagcctaac 2881 tcaaaatact catccaagac agcaggcact ggagcagctg ttggaagaca aggttgaaga 2941 tgatatgctg cagctttcag aatttgaccc cctattgaga gagattgctc ctggccccct 3001 caccacaccc tctgtcccag gctccactcc tggtccctgc ttcctctgtg gttctgcccc 3061 aggcacactg cactgcccat cctgtaaaca ggccctgtgt ccagcctgtg accacctgtt 3121 ccatggacac ccatcccgtg ctcatcacct ccgccagacc ctgcctgggg tcctgcaggg 3181 tacccacctg agccccagtt tacctgcctc agcccaacca cggccccagt cgacctccct 3241 gctggccctg ggagacagct ctctttcttc ccctaatcct gcaagtgctc atttgccctg 3301 gcactgtgct gcctgtgcca tgctaaatga gccttgggca gtgctctgtg tggcctgtga 3361 tcggccccga ggctgtaagg ggttggggtt gggaactgag ggtccccaag gaactggagg 3421 cctagaacct gatcttgcac ggggtcggtg ggcctgccag agctgtacct ttgagaatga 3481 ggcagctgct gtgctatgtt ccatatgtga gcgacctcgg ctggcccagc ctcccagctt 3541 ggtggtggat tcccgagatg ctggcatttg cctgcaaccc cttcagcagg gggatgcttt 3601 gctggcctct gcccagagtc aagtctggta ctgtattcac tgtaccttct gcaactcgag 3661 ccctggctgg gtgtgtgtta tgtgcaaccg gactagtagc cccattccag cacaacatgc 3721 cccccggccc tatgccagct ctttggaaaa gggacccccc aagcctgggc ccccacgacg 3781 ccttagtgcc cccctgccca gttcctgtgg agatcctgag aagcagcgcc aagacaagat 3841 gcgggaagaa ggcctccagc tagtgagcat gatccgggaa ggggaagccg caggtgcctg 3901 tccagaggag atcttctcgg ctctgcagta ctcgggcact gaggtgcctc tgcagtggtt 3961 gcgctcagaa ctgccctacg tcctggagat ggtggctgag ctggctggac agcaggaccc 4021 tgggctgggt gccttttcct gtcaggaggc ccggagagcc tggctggatc gtcatggcaa 4081 ccttgatgaa gctgtggagg agtgtgtgag gaccaggcga aggaaggtgc aggagctcca 4141 gtctctaggc tttgggcctg aggaggggtc tctccaggca ttgttccagc acggaggtga 4201 tgtgtcacgg gccctgactg agctacagcg ccaacgccta gagcccttcc gccagcgcct 4261 ctgggacagt ggccctgagc ccaccccttc ctgggatggg ccagacaagc agagcctggt 4321 caggcggctt ttggcagtct acgcactccc cagctggggc cgggcagagc tggcactgtc 4381 actgctgcag gagacaccca ggaactatga gttgggggat gtggtagaag ctgtgaggca 4441 cagccaggac cgggccttcc tgcgccgctt gcttgcccag gagtgtgccg tgtgtggctg 4501 ggccctgccc cacaaccgga tgcaggccct gacttcctgt gagtgcacca tctgtcctga 4561 ctgcttccgc cagcacttca ccatcgcctt gaaggagaag cacatcacag acatggtgtg 4621 ccctgcctgt ggccgccccg acctcaccga tgacacacag ttgctcagct acttctctac 4681 ccttgacatc cagcttcgcg agagcctaga gccagatgcc tatgcgttgt tccataagaa 4741 gctgaccgag ggtgtgctga tgcgggaccc caagttcttg tggtgtgccc agtgctcctt 4801 tggcttcata tatgagcgtg agcagctgga ggcaacttgt ccccagtgtc accagacctt 4861 ctgtgtgcgc tgcaagcgcc agtgggagga gcagcaccga ggtcggagct gtgaggactt 4921 ccagaactgg aaacgcatga acgacccaga ataccaggcc cagggcctag caatgtatct 4981 tcaggaaaac ggcattgact gccccaaatg caagttctcg tacgccctgg cccgaggagg 5041 ctgcatgcac tttcactgta cccagtgccg ccaccagttc tgcagcggct gctacaatgc 5101 cttttacgcc aagaataaat gtccagagcc taactgcagg gtgaaaaagt ccctgcacgg 5161 ccaccaccct cgagactgcc tcttctacct gcgggactgg actgctctcc ggcttcagaa 5221 gctgctacag gacaataacg tcatgtttaa tacagagcct ccagctgggg cccgggcagt 5281 ccctggaggc ggctgccgag tgatagagca gaaggaggtt cccaatgggc tcagggacga 5341 agcttgtggc aaggaaactc cagctggcta tgccggcctg tgccaggcac actacaaaga 5401 gtatcttgtg agcctcatca atgcccactc gctggaccca gccaccttgt atgaggtgga 5461 agagctggag acggccactg agcgctacct gcacgtacgc ccccagcctt tggctggaga 5521 ggatccccct gcttaccagg cccgcttgtt acagaagctg acagaagagg tacccttggg 5581 acagagtatc ccccgcaggc ggaagggatc ctacccatac gacgtaccag attacgctta 5641 gtctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc 5701 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc 5761 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc 5821 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg 5881 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg 5941 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc 6001 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc 6061 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc 6121 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg 6181 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 6241 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 6301 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 6361 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 6421 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 6481 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 6541 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 6601 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 6661 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 6721 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg 6781 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 6841 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 6901 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 6961 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 7021 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 7081 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 7141 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc 7201 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 7261 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 7321 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 7381 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca 7441 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt 7501 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 7561 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat 7621 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 7681 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 7741 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 7801 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 7861 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 7921 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 7981 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 8041 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 8101 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 8161 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 8221 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 8281 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt 8341 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 8401 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 8461 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 8521 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 8581 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 8641 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 8701 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 8761 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 8821 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 8881 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 8941 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 9001 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 9061 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 9121 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 9181 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 9241 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 9301 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 9361 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 9421 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 9481 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 9541 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 9601 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 9661 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 9721 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 9781 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 9841 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 9901 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 9961 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 10021 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 10081 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 10141 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 10201 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 10261 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 10321 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 10381 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 10441 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 10501 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc 10561 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 10621 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 10681 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 10741 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 10801 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa 10861 acttagtagt //