LOCUS V019990 8564 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019990 VERSION V019990 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..3320 /label="FGF2(NM_001361665)" /note="FGF2(NM_001361665)" /gene="FGF2" promoter 3352..3851 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 3872..4471 /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 4485..5073 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5280..5913 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6042..6058) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6066..6082 /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(6090..6120) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6135..6156 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6444..7032) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7203..8063) /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(8064..8168) /gene="bla" /label="AmpR promoter" polyA_signal 8216..8350 /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 acgctatggc agccgggagc atcaccacgc 2881 tgcccgcctt gcccgaggat ggcggcagcg gcgccttccc gcccggccac ttcaaggacc 2941 ccaagcggct gtactgcaaa aacgggggct tcttcctgcg catccacccc gacggccgag 3001 ttgacggggt ccgggagaag agcgaccctc acatcaagct acaacttcaa gcagaagaga 3061 gaggagttgt gtctatcaaa ggagtgtgtg ctaaccgtta cctggctatg aaggaagatg 3121 gaagattact ggcttctaaa tgtgttacgg atgagtgttt cttttttgaa cgattggaat 3181 ctaataacta caatacttac cggtcaagga aatacaccag ttggtatgtg gcactgaaac 3241 gaactgggca gtataaactt ggatccaaaa caggacctgg gcagaaagct atactttttc 3301 ttccaatgtc tgctaagagc tgaggatccc gcgactctag ataattctac cgggtagggg 3361 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc 3421 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg 3481 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc 3541 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta 3601 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg 3661 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg 3721 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc 3781 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc 3841 cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct 3901 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta 3961 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca 4021 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg 4081 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga 4141 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga 4201 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt 4261 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc 4321 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt 4381 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac 4441 ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt 4501 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 4561 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 4621 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 4681 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 4741 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 4801 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 4861 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 4921 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 4981 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 5041 cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct 5101 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg 5161 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag 5221 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact 5281 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac 5341 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact 5401 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa 5461 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga 5521 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga 5581 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc 5641 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc 5701 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag 5761 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 5821 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 5881 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc 5941 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc 6001 gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 6061 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 6121 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 6181 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 6241 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 6301 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 6361 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 6421 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 6481 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 6541 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 6601 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 6661 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 6721 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 6781 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 6841 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 6901 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 6961 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 7021 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 7081 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 7141 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 7201 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 7261 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 7321 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 7381 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 7441 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 7501 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 7561 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 7621 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 7681 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 7741 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 7801 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 7861 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 7921 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 7981 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 8041 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 8101 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 8161 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt 8221 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 8281 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 8341 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata 8401 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga 8461 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt 8521 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt //