LOCUS V029005 9250 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029005 VERSION V029005 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 1804..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2317..3276 /label="RFC2(NM_002914)" /note="RFC2(NM_002914)" /gene="RFC2" CDS 3283..3348 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3392..3603 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3616..3884 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3913..3922 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3943..4605 /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 4675..4728 /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 4729..5328 /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 5329..5917 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5991..6224 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6296..6417 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6457..6592 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6625..6641) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6649..6665 /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(6673..6703) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6718..6739 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7027..7615) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7786..8646) /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(8647..8751) /gene="bla" /label="AmpR promoter" primer_bind 9225..9241 /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 ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgg aggtggaggc cgtctgtggt 2341 ggcgcgggcg aggtggaggc ccaggactct gaccctgccc ctgccttcag caaggccccc 2401 ggcagcgccg gccactacga actgccgtgg gttgaaaaat ataggccagt aaagctgaat 2461 gaaattgtcg ggaatgaaga caccgtgagc aggctagagg tctttgcaag ggaaggaaat 2521 gtgcccaaca tcatcattgc gggccctcca ggaaccggca agaccacaag cattctgtgc 2581 ttggcccggg ccctgctggg cccagcactc aaagatgcca tgttggaact caatgcttca 2641 aatgacagca tgaccgacgg agcccagcaa gccttgagga gaaccatgga aatctactct 2701 aaaaccactc gcttcgccct tgcttgtaat gcttcggata agatcatcga gcccattcag 2761 tcccgctgtg cagtcctccg gtacacaaag ctgaccgacg cccagatcct caccaggctg 2821 atgaatgtta tcgagaagga gagggtaccc tacactgatg acggcctaga agccatcatc 2881 ttcacggccc agggagacat gaggcaggcg ctgaacaacc tgcagtccac cttctcagga 2941 tttggcttca ttaacagtga gaacgtgttc aaggtctgtg acgagcccca cccactgctg 3001 gtaaaggaga tgatccagca ctgtgtgaat gccaacattg acgaagccta caagattctt 3061 gctcacttgt ggcatctggg ctactcacca gaagatatca ttggcaacat ctttcgagtg 3121 tgtaaaactt tccaaatggc agaatacctg aaactggagt ttatcaagga aattggatac 3181 actcacatga aaatagcgga aggagtgaac tctcttttgc agatggcagg cctcctggca 3241 aggctgtgtc agaagacaat ggccccggtg gccagtggat ccgactacaa agaccatgac 3301 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 3361 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 3421 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 3481 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 3541 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 3601 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 3661 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 3721 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 3781 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 3841 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 3901 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 3961 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 4021 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 4081 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 4141 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 4201 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 4261 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4321 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 4381 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 4441 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 4501 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 4561 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 4621 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 4681 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 4741 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 4801 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 4861 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 4921 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 4981 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5041 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5101 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5161 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5221 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5281 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 5341 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5401 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5461 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 5521 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5581 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 5641 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 5701 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 5761 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 5821 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 5881 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 5941 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 6001 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 6061 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6121 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6181 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6241 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6301 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6361 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6421 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 6481 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 6541 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 6601 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 6661 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6721 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6781 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6841 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6901 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6961 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7021 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7081 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7141 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7201 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7261 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7321 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7381 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7441 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7501 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7561 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7621 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7681 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7741 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7801 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7861 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7921 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7981 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8041 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8101 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8161 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8221 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8281 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8341 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8401 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8461 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8521 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8581 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8641 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8701 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8761 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8821 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8881 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8941 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9001 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9061 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9121 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9181 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9241 tgccaagctg //