LOCUS V018352 8995 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018352 VERSION V018352 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..3751 /label="Cdk4(NM_009870)" /note="Cdk4(NM_009870)" /gene="Cdk4" promoter 3783..4282 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4303..4902 /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 4916..5504 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5711..6344 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6473..6489) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6497..6513 /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(6521..6551) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6566..6587 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6875..7463) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7634..8494) /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(8495..8599) /gene="bla" /label="AmpR promoter" polyA_signal 8647..8781 /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 ccatggctgc cactcgatat gaacccgtgg ctgaaattgg 2881 tgtcggtgcc tatgggacgg tgtacaaagc ccgagatccc cacagtggcc actttgtggc 2941 cctcaagagt gtgagagttc ctaatggagg agcagctgga gggggccttc ccgtcagcac 3001 agttcgtgag gtggccttgt taaggaggct ggaggccttt gaacatccca atgttgtacg 3061 gctgatggat gtctgtgcta cttcccgaac tgatcgggac atcaaggtca ccctagtgtt 3121 tgagcatata gaccaggacc tgaggacata cctggacaaa gcacctccac cgggcctgcc 3181 ggttgagacc attaaggatc taatgcgtca gtttctaagc ggcctggatt ttcttcatgc 3241 aaactgcatt gttcaccggg acctgaagcc agagaacatt ctagtgacaa gtaatgggac 3301 cgtcaagctg gctgactttg gcctagctag aatctacagc taccagatgg ccctcacgcc 3361 tgtggtggtt acgctctggt accgagctcc tgaagttctt ctgcagtcta catacgcaac 3421 acccgtggac atgtggagcg ttggctgtat ctttgcagag atgttccgtc ggaagcctct 3481 cttctgtgga aactctgaag ccgaccagtt ggggaaaatc tttgatctca ttggattgcc 3541 tccagaagac gactggcctc gagaggtatc tctacctcga ggagcctttg cccccagagg 3601 gcctcggcca gtgcagtcag tggtgccaga gatggaggag tctggagcgc agctgctact 3661 ggaaatgctg acctttaacc cacataagcg aatctctgcc ttccgagccc tgcagcactc 3721 ctacctgcac aaggaggaaa gcgacgcaga gtgaggatcc cgcgactcta gataattcta 3781 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct 3841 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag 3901 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt 3961 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc 4021 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct 4081 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag 4141 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc 4201 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc 4261 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc 4321 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 4381 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 4441 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 4501 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 4561 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 4621 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 4681 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 4741 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 4801 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 4861 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca 4921 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 4981 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 5041 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 5101 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 5161 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 5221 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 5281 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 5341 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 5401 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 5461 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc 5521 agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct 5581 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag 5641 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa 5701 aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg 5761 atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc 5821 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta 5881 gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg 5941 gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac 6001 gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag 6061 ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga 6121 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga 6181 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6241 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6301 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6361 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg 6421 acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 6481 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 6541 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 6601 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 6661 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 6721 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 6781 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 6841 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 6901 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 6961 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 7021 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 7081 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 7141 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 7201 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 7261 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 7321 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 7381 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 7441 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 7501 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 7561 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 7621 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 7681 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 7741 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 7801 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 7861 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 7921 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 7981 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 8041 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 8101 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 8161 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 8221 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 8281 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 8341 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 8401 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 8461 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 8521 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 8581 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 8641 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 8701 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 8761 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt 8821 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt 8881 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac 8941 ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //