LOCUS V035934 10006 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035934 VERSION V035934 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..3559 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EGFP" CDS 3629..4681 /label="ATF4(NM_182810)" /note="ATF4(NM_182810)" /gene="ATF4" CDS 4688..4714 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4718..4744 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4748..4774 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 4794..5293 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5314..5913 /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 5927..6515 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6722..7355 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7484..7500) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7508..7524 /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(7532..7562) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7577..7598 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7886..8474) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8645..9505) /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(9506..9610) /gene="bla" /label="AmpR promoter" polyA_signal 9658..9792 /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 ccatggtgag caagggcgag gagctgttca ccggggtggt 2881 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 2941 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 3001 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 3061 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 3121 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 3181 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 3241 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 3301 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 3361 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 3421 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 3481 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 3541 catggacgag ctgtacaagt ccggactcag atctcgaggc ggcggaggga gtggaggagg 3601 gggaagcgga ggcggaggga gtggatccat gaccgaaatg agcttcctga gcagcgaggt 3661 gttggtgggg gacttgatgt cccccttcga ccagtcgggt ttgggggctg aagaaagcct 3721 aggtctctta gatgattacc tggaggtggc caagcacttc aaacctcatg ggttctccag 3781 cgacaaggct aaggcgggct cctccgaatg gctggctgtg gatgggttgg tcagtccctc 3841 caacaacagc aaggaggatg ccttctccgg gacagattgg atgttggaga aaatggattt 3901 gaaggagttc gacttggatg ccctgttggg tatagatgac ctggaaacca tgccagatga 3961 ccttctgacc acgttggatg acacttgtga tctctttgcc cccctagtcc aggagactaa 4021 taagcagccc ccccagacgg tgaacccaat tggccatctc ccagaaagtt taacaaaacc 4081 cgaccaggtt gcccccttca ccttcttaca acctcttccc ctttccccag gggtcctgtc 4141 ctccactcca gatcattcct ttagtttaga gctgggcagt gaagtggata tcactgaagg 4201 agataggaag ccagactaca ctgcttacgt tgccatgatc cctcagtgca taaaggagga 4261 agacacccct tcagataatg atagtggcat ctgtatgagc ccagagtcct atctggggtc 4321 tcctcagcac agcccctcta ccaggggctc tccaaatagg agcctcccat ctccaggtgt 4381 tctctgtggg tctgcccgtc ccaaacctta cgatcctcct ggagagaaga tggtagcagc 4441 aaaagtaaag ggtgagaaac tggataagaa gctgaaaaaa atggagcaaa acaagacagc 4501 agccactagg taccgccaga agaagagggc ggagcaggag gctcttactg gtgagtgcaa 4561 agagctggaa aagaagaacg aggctctaaa agagagggcg gattccctgg ccaaggagat 4621 ccagtacctg aaagatttga tagaagaggt ccgcaaggca agggggaaga aaagggtccc 4681 cggatcctac ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta 4741 tgctagctat ccttatgatg tcccagacta cgcatagtct agataattct accgggtagg 4801 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg 4861 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc 4921 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt 4981 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac 5041 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag 5101 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc 5161 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg 5221 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc 5281 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc 5341 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac 5401 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg 5461 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac 5521 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc 5581 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg 5641 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc 5701 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag 5761 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc 5821 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc 5881 acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga 5941 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 6001 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 6061 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 6121 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 6181 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 6241 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 6301 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac 6361 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 6421 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 6481 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac 6541 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc 6601 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta 6661 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga 6721 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac 6781 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca 6841 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc 6901 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg 6961 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga 7021 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc 7081 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga 7141 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg 7201 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 7261 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 7321 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat 7381 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta 7441 gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 7501 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 7561 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 7621 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 7681 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 7741 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 7801 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 7861 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 7921 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 7981 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 8041 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 8101 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 8161 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 8221 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 8281 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 8341 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 8401 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 8461 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 8521 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 8581 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 8641 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 8701 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 8761 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 8821 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 8881 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 8941 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 9001 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 9061 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 9121 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 9181 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 9241 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 9301 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 9361 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 9421 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 9481 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 9541 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 9601 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac 9661 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 9721 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 9781 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca 9841 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct 9901 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt 9961 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt //