LOCUS V018354 10959 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018354 VERSION V018354 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 2827..5715 /label="Thbs4(NM_011582)" /note="Thbs4(NM_011582)" /gene="Thbs4" promoter 5747..6246 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 6267..6866 /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 6880..7468 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7675..8308 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8437..8453) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8461..8477 /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(8485..8515) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8530..8551 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8839..9427) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9598..10458) /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(10459..10563) /gene="bla" /label="AmpR promoter" polyA_signal 10611..10745 /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 gccaccatgc cggccccacg cgcggccgcg gccgccttcc tcctgctgca cctggtcctg 2881 cagccgtggc agcggacgag cgcccaggcc acaccccagg tctttgatct tctaccgtcc 2941 tccagccaga ggctgaaccc cagcgccctg cagccagtcc tgacagatcc caccctgcac 3001 gaagtctatc tcatctccac cttcaagctg cagagtaaaa gctcagccac catctttggc 3061 ctctactcct caagtgacaa cagcaaatac tttgaattca ctgtgatggg acggctgaac 3121 aaagccatcc tccgctacct gaagaatgat ggaaagatcc atctggtggt gttcaacaac 3181 ttgcagctgg ctgatgggag gcgccacagg gttctcctga gactgagcaa tttgcagaga 3241 ggagatggct ctgtagaact ctacctggac tgcgctcagg cggactcggt gcgcaacctc 3301 cccagagcct tttctggcct cacccagaac ccagagtcca ttgaattgcg gacattccag 3361 aggaagcccc aggacttctt ggaagagctg aagctggtgg tgcggggctc gctgttccag 3421 gtggccagcc tccaagactg cttcctgcag cagagtgagc ctctggctgc cacaagcaca 3481 ggagacttta atcgacagtt cttgggtcaa atgacacaac tgaaccagct ccttggagag 3541 gtgaaagatc ttctgaggca gcaggtcaag gaaacatcgt ttttgcgaaa caccattgct 3601 gaatgccaag cttgtggtcc tctcagcttt cagtccccaa ctccaaacac actggtgccc 3661 atagctcctc cagcaccccc aactcgccca acacgccact gtgactccag cccttgtttc 3721 cgaggtgtcc gatgtacgga caccagggat ggcttccagt gtggaccctg cccagatggt 3781 tacacgggaa acgggatcac ctgctctgac gtggatgagt gcaaatacca tccctgctat 3841 ccaggtgtgc gctgtgtgaa tttggctccg ggtttcagat gtgacgcctg tccagtaggt 3901 ttcacagggc ccatggtgca gggtgtcggg atcaactttg ctaaaacaaa caagcaggtg 3961 tgcactgatg ttgatgagtg tcagaatgga gcgtgtgttc tgaattctat ctgcattaac 4021 actttgggat cttatcgctg tgggccttgc aaaccagggt acactggtga tcagacgagg 4081 ggatgcaaga ctgaaaggag ctgccggaac ccagagcaga acccatgcag tgtgcatgcg 4141 cagtgcattg aggagcggca aggggatgtg acgtgtgtgt gtggtgttgg ctgggctggt 4201 gacggctatg tctgtggaaa ggatgtggac attgacagtt accctgatga agaactgcca 4261 tgctctgcca ggaactgcaa gaaggataac tgcaagtatg tgccaaactc tggccaggaa 4321 gatgcagaca gagatggcat cggagacgcg tgtgatgagg atgcagatgg agatgggatc 4381 ttgaatgaac aggacaactg tgtcctgact cacaatatag accaaaggaa cagtgataaa 4441 gatatctttg gggatgcctg tgacaactgc cggatggtcc tgaataatga ccagaaggat 4501 actgacgggg atgggagagg agacgcctgt gatgacgaca tggatggaga tggaataaaa 4561 aacattttgg acaactgtcc cagagttccc aaccgtgacc agcaggatcg ggatggtgat 4621 gatgtcggag atgcttgtga cagctgtcct gatgtcagta accctaacca gtcggatgtg 4681 gataacgatc tggttgggga ctcctgtgac accaaccaag acagtgacgg ggatggccac 4741 caggacagca cagacaactg ccccacagtc attaacagct cccagctgga cactgacaaa 4801 gatgggatcg gcgatgaatg tgatgacgat gacgataacg atggcatccc agacctggtg 4861 cctcctggcc cagacaactg caggctcgtc cccaacccag cccaggaaga tagcaacaat 4921 gacggggtag gagatatctg tgaagctgac ttcgaccagg accaggtcat cgatcacatt 4981 gatgtgtgcc ctgagaatgc ggagatcacc ctgactgact tcagggccta ccagacagtg 5041 gtcctggacc ctgaagggga tgctcagatc gaccccaact gggtggtcct gaaccagggc 5101 atggaaattg tgcagaccat gaacagtgac cctggcctag ctgttgggta cacggcattt 5161 aacggagttg actttgaagg gaccttccac gtaaacaccc agacagatga tgactatgct 5221 ggctttattt ttggttacca agacagctcc agcttctatg tagtcatgtg gaagcaaacg 5281 gagcagacat actggcaggc tacaccgttc cgtgcagtag cagaacctgg tattcagctc 5341 aaggctgtga agtctaaaac tggcccaggg gaacatctcc gaaactccct gtggcacaca 5401 ggggacacca gcgaccaggt caggctgctg tggaaggact ccaggaatgt gggctggaag 5461 gacaaggtct cctaccgctg gttcctgcag cataggcccc aggtgggcta catcagggtg 5521 cgcttttatg aaggctcgga gttggtggct gactctgggg tcaccataga caccaccatg 5581 cgtggaggcc gacttggtgt gttctgcttc tctcaagaaa acatcatctg gtccaacctc 5641 aagtaccgct gcaacgacac cataccagag gacttccagg agttccagac ccagagtttc 5701 gaccgcttgg ataattgagg atcccgcgac tctagataat tctaccgggt aggggaggcg 5761 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac 5821 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg 5881 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc 5941 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc 6001 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa 6061 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc 6121 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca 6181 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc 6241 ctttcgacct gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca 6301 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg 6361 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac 6421 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg 6481 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg 6541 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc 6601 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc 6661 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg 6721 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg 6781 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt 6841 gcatgacccg caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa 6901 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 6961 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 7021 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 7081 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 7141 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 7201 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 7261 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 7321 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 7381 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 7441 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa 7501 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag 7561 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa 7621 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag 7681 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag 7741 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct 7801 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag 7861 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag 7921 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc 7981 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg 8041 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca 8101 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg 8161 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 8221 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 8281 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat 8341 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgtttt 8401 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 8461 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 8521 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 8581 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 8641 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 8701 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 8761 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 8821 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 8881 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 8941 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 9001 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 9061 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 9121 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 9181 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 9241 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 9301 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 9361 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 9421 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 9481 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 9541 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 9601 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 9661 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 9721 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 9781 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 9841 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 9901 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 9961 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 10021 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 10081 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 10141 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 10201 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 10261 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 10321 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 10381 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 10441 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 10501 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 10561 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta 10621 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 10681 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 10741 ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta 10801 ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat 10861 tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta gtttttaaag 10921 aaattgtatt tgttaaatat gtactacaaa cttagtagt //