LOCUS V029663 9373 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029663 VERSION V029663 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..3399 /label="SURF6(NM_006753)" /note="SURF6(NM_006753)" /gene="SURF6" CDS 3406..3471 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3515..3726 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3739..4007 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4036..4045 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4066..4728 /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 4798..4851 /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 4852..5451 /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 5452..6040 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6114..6347 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6419..6540 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6580..6715 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6748..6764) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6772..6788 /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(6796..6826) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6841..6862 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7150..7738) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7909..8769) /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(8770..8874) /gene="bla" /label="AmpR promoter" primer_bind 9348..9364 /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 cctctctact cgccaaggac 2341 gcctacctgc agagcctggc caagaagatc tgctcccatt cggccccgga acagcaggcg 2401 cgcacgcggg ctggcaaaac tcaaggctca gaaactgcag ggcccccaaa aaagaaaagg 2461 aagaaaacac aaaagaaatt ccggaagcga gaagagaagg ctgctgagca caaggccaag 2521 tccttggggg agaaatctcc agcagcctct ggggccagga ggcctgaggc agccaaagag 2581 gaagcagctt gggcttccag ctcagcaggg aaccctgcag atggcctggc cactgagcct 2641 gagtctgtct ttgctctgga tgttctgcga cagcgactgc atgagaagat ccaggaggcc 2701 cggggccagg gcagtgccaa ggagctgtcc cctgccgcct tggagaaaag gcggcggaga 2761 aagcaggaac gggaccggaa gaagaggaag cgaaaggagc tgcgggcgaa agagaaggcc 2821 aggaaggctg aggaggccac ggaggcccag gaggtggtgg aggcaacccc agagggggcc 2881 tgcacggagc cgcgggagcc gcccgggctg atcttcaata aggtggaggt gagcgaagac 2941 gagccggcca gcaaggcgca gcgcagaaaa gagaagaggc agagggtgaa ggggaacctc 3001 acgccgctga ccgggaggaa ctaccggcag ctgctggagc gcctgcaggc acggcagagc 3061 cggctggacg agctgcgcgg ccaggatgag gggaaggcgc aggagctgga ggcgaagatg 3121 aagtggacca acctcctcta caaggcggag ggcgtgaaga tccgtgacga cgaacgcctg 3181 ctgcaggagg ccctgaagcg caaggagaag cgcagggcgc agcggcagcg ccggtgggag 3241 aagcgcacgg ccggcgtggt ggagaagatg cagcagcgcc aggaccggcg gcggcagaac 3301 ctgcgcagga agaaggcggc ccgcgccgag cgccgcctgc tcagagcccg caagaagggc 3361 cgcatcctgc cgcaggacct ggagcgcgca ggcctggtcg gatccgacta caaagaccat 3421 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3481 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 3541 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3601 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 3661 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 3721 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 3781 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 3841 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 3901 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 3961 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4021 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 4081 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 4141 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 4201 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 4261 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 4321 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 4381 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 4441 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 4501 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 4561 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 4621 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 4681 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 4741 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 4801 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 4861 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4921 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4981 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 5041 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 5101 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 5161 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 5221 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 5281 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 5341 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5401 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 5461 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5521 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5581 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5641 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5701 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5761 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5821 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 5881 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5941 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6001 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 6061 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 6121 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 6181 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6241 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6301 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6361 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6421 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6481 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 6541 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 6601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 6661 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 6721 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 6781 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6841 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6901 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6961 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7021 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7081 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7141 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7201 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7261 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7321 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7381 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7441 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7501 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7561 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7621 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7681 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7741 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7801 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7861 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7921 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7981 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8041 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8101 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8161 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8221 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8281 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8341 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8401 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8461 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8521 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8581 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8641 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8701 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8761 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8821 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8881 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8941 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9001 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9061 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9121 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9181 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9241 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9301 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9361 cagtgccaag ctg //