LOCUS V038422 9664 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038422 VERSION V038422 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" regulatory 2279..2288 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2288..2359 /codon_start=1 /product="three tandem FLAG® epitope tags" /transl_table=1 /translation="DYKDDDDKDYKDDDDKDYKDDDDK" /label="3xFLAG" CDS 2366..3772 /label="MMP1(NM_002421)" /note="MMP1(NM_002421)" /gene="MMP1" promoter 3823..4034 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4047..4315 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4344..4353 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4374..5036 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 5106..5159 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 5160..5759 /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 5760..6347 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6421..6654 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6726..6847 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6866..7001 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7039..7055) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7063..7079 /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(7087..7117) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7132..7153 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7441..8029) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8200..9060) /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(9061..9165) /gene="bla" /label="AmpR promoter" primer_bind 9639..9655 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 caccatggac tacaaggatg acgatgacaa ggattacaaa gacgacgatg ataaggacta 2341 taaggatgat gacgacaaag aattcatgca cagctttcct ccactgctgc tgctgctgtt 2401 ctggggtgtg gtgtctcaca gcttcccagc gactctagaa acacaagagc aagatgtgga 2461 cttagtccag aaatacctgg aaaaatacta caacctgaag aatgatggga ggcaagttga 2521 aaagcggaga aatagtggcc cagtggttga aaaattgaag caaatgcagg aattctttgg 2581 gctgaaagtg actgggaaac cagatgctga aaccctgaag gtgatgaagc agcccagatg 2641 tggagtgcct gatgtggctc agtttgtcct cactgagggg aaccctcgct gggagcaaac 2701 acatctgacc tacaggattg aaaattacac gccagatttg ccaagagcag atgtggacca 2761 tgccattgag aaagccttcc aactctggag taatgtcaca cctctgacat tcaccaaggt 2821 ctctgagggt caagcagaca tcatgatatc ttttgtcagg ggagatcatc gggacaactc 2881 tccttttgat ggacctggag gaaatcttgc tcatgctttt caaccaggcc caggtattgg 2941 aggggatgct cattttgatg aagatgaaag gtggaccaac aatttcagag agtacaactt 3001 acatcgtgtt gcagctcatg aactcggcca ttctcttgga ctctcccatt ctactgatat 3061 cggggctttg atgtacccta gctacacctt cagtggtgat gttcagctag ctcaggatga 3121 cattgatggc atccaagcca tatatggacg ttcccaaaat cctgtccagc ccatcggccc 3181 acaaacccca aaagcgtgtg acagtaagct aacctttgat gctataacta cgattcgggg 3241 agaagtgatg ttctttaaag acagattcta catgcgcaca aatcccttct acccggaagt 3301 tgagctcaat ttcatttctg ttttctggcc acaactgcca aatgggcttg aagctgctta 3361 cgaatttgcc gacagagatg aagtccggtt tttcaaaggg aataagtact gggctgttca 3421 gggacagaat gtgctacacg gataccccaa ggacatctac agctcctttg gcttccctag 3481 aactgtgaag catatcgatg ctgctctttc tgaggaaaac actggaaaaa cctacttctt 3541 tgttgctaac aaatactgga ggtatgatga atataaacga tctatggatc caggttatcc 3601 caaaatgata gcacatgact ttcctggaat tggccacaaa gttgatgcag ttttcatgaa 3661 agatggattt ttctatttct ttcatggaac aagacaatac aaatttgatc ctaaaacgaa 3721 gagaattttg actctccaga aagctaatag ctggttcaac tgcaggaaaa attgaggatc 3781 cgcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc 3841 ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt 3901 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg 3961 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg 4021 ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc 4081 ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg 4141 cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt 4201 gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc 4261 tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc 4321 tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc ctgcccgcca 4381 tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag ctggtgggcg 4441 gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc accaaaggcg 4501 ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc taccacttcg 4561 gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac ggcggctaca 4621 ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc 4681 gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc ttccccgagg 4741 acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag cacctgcacc 4801 ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg 4861 gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc caccccagca 4921 tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg cacagcaaca 4981 ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc ttcgccagat 5041 cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc tccaccggat 5101 ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta 5161 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac 5221 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc 5281 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca 5341 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga 5401 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt 5461 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc 5521 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca 5581 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg 5641 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg 5701 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa 5761 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 5821 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 5881 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 5941 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 6001 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 6061 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 6121 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg 6181 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 6241 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc 6301 gccttcgccc tcagacgagt cggatctccc tttggccgcc tccccgcctg gtacctttaa 6361 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 6421 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 6481 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 6541 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 6601 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 6661 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6721 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6781 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6841 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6901 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6961 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 7021 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 7081 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 7141 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7201 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7261 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7321 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7381 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7441 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7501 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7561 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7621 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7681 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7741 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7801 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7861 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7921 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7981 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 8041 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 8101 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 8161 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8221 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8281 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8341 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8401 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8461 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8521 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8581 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8641 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8701 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8761 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8821 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8881 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8941 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 9001 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 9061 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 9121 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9181 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9241 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9301 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9361 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9421 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9481 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9541 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9601 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9661 gctg //