LOCUS V021559 8636 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021559 VERSION V021559 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 2826..3392 /label="HRAS(NM_005343)" /note="HRAS(NM_005343)" /gene="HRAS" promoter 3424..3923 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 3944..4543 /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 4557..5145 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5352..5985 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6114..6130) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6138..6154 /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(6162..6192) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6207..6228 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6516..7104) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7275..8135) /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(8136..8240) /gene="bla" /label="AmpR promoter" polyA_signal 8288..8422 /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 ccaccatgac cgaatacaag cttgttgttg ttggcgccgt cggtgtgggc aagagtgcgc 2881 tgaccatcca gctgatccag aaccattttg tggacgaata cgaccccact atagaggatt 2941 cctaccggaa gcaggtggtc attgatgggg agacgtgcct gttggacatc ctggataccg 3001 ccggccagga ggagtacagc gccatgcggg accagtacat gcgcaccggg gagggcttcc 3061 tgtgtgtgtt tgccatcaac aacaccaagt cttttgagga catccaccag tacagggagc 3121 agatcaaacg ggtgaaggac tcggatgacg tgcccatggt gctggtgggg aacaagtgtg 3181 acctggctgc acgcactgtg gaatctcggc aggctcagga cctcgcccga agctacggca 3241 tcccctacat cgagacctcg gccaagaccc ggcagggagt ggaggatgcc ttctacacgt 3301 tggtgcgtga gatccggcag cacaagctgc ggaagctgaa ccctcctgat gagagtggcc 3361 ccggctgcat gagctgcaag tgtgtgctct cctgaggatc ccgcgactct agataattct 3421 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc 3481 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta 3541 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag 3601 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag 3661 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc 3721 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa 3781 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt 3841 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct 3901 cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc 3961 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc 4021 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt 4081 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 4141 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 4201 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 4261 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 4321 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 4381 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 4441 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga 4501 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc 4561 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 4621 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 4681 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 4741 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 4801 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 4861 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 4921 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 4981 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 5041 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 5101 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg 5161 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc 5221 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca 5281 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 5341 aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg 5401 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt 5461 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt 5521 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat 5581 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca 5641 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa 5701 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg 5761 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag 5821 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat 5881 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact 5941 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc 6001 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt 6061 gacattgcta gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag 6121 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 6181 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 6241 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 6301 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 6361 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 6421 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 6481 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 6541 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 6601 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 6661 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 6721 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 6781 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 6841 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 6901 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 6961 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 7021 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 7081 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 7141 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 7201 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 7261 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 7321 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 7381 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 7441 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 7501 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 7561 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 7621 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 7681 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 7741 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 7801 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 7861 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 7921 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 7981 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 8041 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 8101 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 8161 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 8221 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg 8281 ggagatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 8341 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 8401 tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact 8461 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg 8521 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa 8581 cttagtagtt tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt //