LOCUS V021227 9010 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021227 VERSION V021227 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..3767 /label="NHEJ1(NM_001377499)" /note="NHEJ1(NM_001377499)" /gene="NHEJ1" promoter 3798..4297 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4318..4917 /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 4931..5519 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5726..6359 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6488..6504) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6512..6528 /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(6536..6566) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6581..6602 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6890..7478) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7649..8509) /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(8510..8614) /gene="bla" /label="AmpR promoter" polyA_signal 8662..8796 /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 agaactggag caaggcctgt 2881 tgatgcagcc atgggcgtgg ctacagcttg cagagaactc cctcttggcc aaggttttta 2941 tcaccaagca gggctatgcc ttgttggttt cagatcttca acaggtgtgg catgaacagg 3001 tggacactag tgtggtcagc cagcgagcca aggagctgaa caagcggctc actgctcctc 3061 ctgcagcttt cctctgtcat ttggataatc tccttcgccc attgttgaag gacgctgctc 3121 accctagcga agctaccttc tcctgtgatt gtgtggcaga tgcactgatt ctacgggtgc 3181 gaagtgagct ctctggcctc cccttctatt ggaatttcca ctgcatgcta gctagtcctt 3241 ccctggtctc ccaacatttg attcgtcctc tgatgggcat gagtctggca ttacagtgcc 3301 aagtgaggga gctagcaacg ttacttcata tgaaagacct agagatccaa gactaccagg 3361 agagtggggc tacgctgatt cgagatcgat tgaagacaga accatttgaa gaaaattcct 3421 tcttggaaca atttatgata gagaaactgc cagaggcatg cagcattggt gatggaaagc 3481 cctttgtcat gaatctgcag gatctgtata tggcagtcac cacacaagag gtccaagtgg 3541 gacagaagca tcaaggcgct ggtgagccct ccagaggaga tcctcatacc tcaaacagtg 3601 cttccctgca aggaatcgat agccaatgtg taaaccagcc agaacaactg gtctcctcag 3661 ccccaaccct ctcagcacct gagaaagagt ccacgggtac ttcaggccct ctgcagagac 3721 ctcagctgtc aaaggtcaag aggaagaagc caaggggtct cttcagttag gatcccgcga 3781 ctctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc 3841 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc 3901 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc 3961 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg 4021 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg 4081 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc 4141 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc 4201 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc 4261 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg 4321 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 4381 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 4441 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 4501 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 4561 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 4621 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 4681 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 4741 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 4801 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 4861 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg 4921 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 4981 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 5041 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 5101 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 5161 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 5221 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 5281 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc 5341 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 5401 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 5461 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 5521 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca 5581 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt 5641 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 5701 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat 5761 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 5821 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 5881 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 5941 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 6001 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 6061 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 6121 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 6181 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 6241 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 6301 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 6361 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 6421 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt 6481 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 6541 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 6601 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 6661 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 6721 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 6781 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 6841 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 6901 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 6961 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 7021 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 7081 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 7141 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 7201 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 7261 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 7321 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 7381 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 7441 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 7501 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 7561 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 7621 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 7681 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 7741 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 7801 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 7861 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 7921 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 7981 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 8041 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 8101 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 8161 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 8221 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 8281 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 8341 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 8401 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 8461 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 8521 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 8581 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 8641 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc 8701 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 8761 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 8821 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 8881 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 8941 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa 9001 acttagtagt //