LOCUS V035740 9329 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035740 VERSION V035740 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..4085 /label="CCNE1(NM_001238)" /note="CCNE1(NM_001238)" /gene="CCNE1" promoter 4117..4616 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4637..5236 /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 5250..5838 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6045..6678 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6807..6823) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6831..6847 /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(6855..6885) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6900..6921 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7209..7797) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7968..8828) /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(8829..8933) /gene="bla" /label="AmpR promoter" polyA_signal 8981..9115 /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 acgctatgcc gagggagcgc agggagcggg 2881 atgcgaagga gcgggacacc atgaaggagg acggcggcgc ggagttctcg gctcgctcca 2941 ggaagaggaa ggcaaacgtg accgtttttt tgcaggatcc agatgaagaa atggccaaaa 3001 tcgacaggac ggcgagggac cagtgtggga gccagccttg ggacaataat gcagtctgtg 3061 cagacccctg ctccctgatc cccacacctg acaaagaaga tgatgaccgg gtttacccaa 3121 actcaacgtg caagcctcgg attattgcac catccagagg ctccccgctg cctgtactga 3181 gctgggcaaa tagagaggaa gtctggaaaa tcatgttaaa caaggaaaag acatacttaa 3241 gggatcagca ctttcttgag caacaccctc ttctgcagcc aaaaatgcga gcaattcttc 3301 tggattggtt aatggaggtg tgtgaagtct ataaacttca cagggagacc ttttacttgg 3361 cacaagattt ctttgaccgg tatatggcga cacaagaaaa tgttgtaaaa actcttttac 3421 agcttattgg gatttcatct ttatttattg cagccaaact tgaggaaatc tatcctccaa 3481 agttgcacca gtttgcgtat gtgacagatg gagcttgttc aggagatgaa attctcacca 3541 tggaattaat gattatgaag gcccttaagt ggcgtttaag tcccctgact attgtgtcct 3601 ggctgaatgt atacatgcag gttgcatatc taaatgactt acatgaagtg ctactgccgc 3661 agtatcccca gcaaatcttt atacagattg cagagctgtt ggatctctgt gtcctggatg 3721 ttgactgcct tgaatttcct tatggtatac ttgctgcttc ggccttgtat catttctcgt 3781 catctgaatt gatgcaaaag gtttcagggt atcagtggtg cgacatagag aactgtgtca 3841 agtggatggt tccatttgcc atggttataa gggagacggg gagctcaaaa ctgaagcact 3901 tcaggggcgt cgctgatgaa gatgcacaca acatacagac ccacagagac agcttggatt 3961 tgctggacaa agcccgagca aagaaagcca tgttgtctga acaaaatagg gcttctcctc 4021 tccccagtgg gctcctcacc ccgccacaga gcggtaagaa gcagagcagc gggccggaaa 4081 tggcgtgagg atcccgcgac tctagataat tctaccgggt aggggaggcg cttttcccaa 4141 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc 4201 tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg 4261 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc 4321 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg 4381 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt 4441 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg 4501 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc 4561 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct 4621 gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga 4681 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca 4741 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac 4801 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt 4861 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat 4921 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc 4981 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca 5041 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg 5101 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg 5161 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg 5221 caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa 5281 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 5341 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 5401 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 5461 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 5521 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 5581 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 5641 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga 5701 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 5761 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 5821 tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc 5881 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga 5941 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa 6001 ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca 6061 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc 6121 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg 6181 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac 6241 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga 6301 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta 6361 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg 6421 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc 6481 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg 6541 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag 6601 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag 6661 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg 6721 caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgtttt accgtcgacc 6781 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 6841 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 6901 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 6961 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 7021 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 7081 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 7141 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 7201 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 7261 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 7321 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 7381 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 7441 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 7501 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 7561 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 7621 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 7681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 7741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 7801 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 7861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 7921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 7981 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 8041 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 8101 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 8161 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 8221 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 8281 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 8341 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 8401 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 8461 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 8521 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 8581 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 8641 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 8701 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 8761 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 8821 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 8881 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 8941 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta 9001 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 9061 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg 9121 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc 9181 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta 9241 aagaaattgt atttgttaaa tatgtactac aaacttagta gtttttaaag aaattgtatt 9301 tgttaaatat gtactacaaa cttagtagt //