LOCUS V038371 9733 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038371 VERSION V038371 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..3759 /label="Fut10(NM_001012517)" /note="Fut10(NM_001012517)" /gene="Fut10" CDS 3766..3831 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3875..4086 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4099..4367 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4396..4405 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4426..5088 /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 5158..5211 /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 5212..5811 /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 5812..6400 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6474..6707 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6779..6900 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6940..7075 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7108..7124) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7132..7148 /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(7156..7186) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7201..7222 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7510..8098) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8269..9129) /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(9130..9234) /gene="bla" /label="AmpR promoter" primer_bind 9708..9724 /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 tgcggtttca gaggaggaag 2341 cttctggcct cttgcctgtg tgtgacagcg actgtctttc tgatggtcac actccaggta 2401 gtggttgagc ttgggaaatt tgaaaggaag aagcttaaag actccaatgt gcaagacgga 2461 catagagatg tggagggaga gcctaaacac ctggagccgt ttcctgaaaa ggaagcactg 2521 gccttggctg ggaggacaaa ggtggatgct ggtagctacc ccatcgtgct gtggtggtcc 2581 ccactgactg gagagactgg caggcttggc cagtgtgggg cagatgcttg cttctttacc 2641 atcaacagga ccttccagca tcatccgatg accagggcct tcctattcta cggtactgac 2701 tttaacatag acagcttacc tttgcctcgg gaagcccatc atgactgggc gcttttccat 2761 gaagagtccc ccaaaaacaa ttataagctc ttccataaac cagtcatcac cttattcaac 2821 catacggcca cattcagcag gcattcccac ttgccactga ctacgcagta cctggaaggc 2881 gtggacgtcc tgaagtcact cagataccta gttcctttac aggctaagaa caacctaaga 2941 caaaaactgg ccccattggt atatgttcag tcggactgcg atccaccatc agacagggac 3001 agctatgtcc gggagctgat ggcgtacatt gaagttgatt cctatggcga gtgtttacag 3061 aacagagatc ttccccagca gctgaaaaac cccgcctcca tggatgctga tgctttctac 3121 agagtcattg cccagtataa gttcatcctg gcctttgaga atgcggtctg tgacgattac 3181 atcacagaga agttctggag accactgaaa ctgggggtcg tgcccgtgta ttacggatcc 3241 cccaccatcg ccgactggct tccaagtaac cgaagcgcca ttctggtgtc ggaattctct 3301 caccccagag agctggcgag cttcattagg agactggatt atgacgatgg gttgtatgag 3361 acctatgtag agtggaagct aaagggcaag atctctaacc agcggcttct gacagctctc 3421 aatgaacggg agtggggtgt gcaggacatt aaccaggaca actacattga ctcgtttgag 3481 tgtatggtgt gccgcagggt gtgggcaaac agtaggctgc aggagcaggg cttaccaccc 3541 aaacaatgga aggcagatgt tagccacctg cattgcccgg agcctgcctt gttcaccttc 3601 tcatctccag cttctccggc cctacgtggg agatcgttgc gagagctgtg gctacctagc 3661 ttccaacagt ctaaaaaaga agcccaggca ctgaggtggc tggtggatag gaatcaaaat 3721 ttttcatctg aagagttctg ggctctagta tttaaggacg gatccgacta caaagaccat 3781 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3841 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 3901 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3961 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 4021 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 4081 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 4141 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 4201 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 4261 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 4321 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4381 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 4441 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 4501 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 4561 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 4621 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 4681 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 4741 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 4801 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 4861 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 4921 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 4981 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 5041 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 5101 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 5161 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 5221 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 5281 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 5341 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 5401 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 5461 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 5521 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 5581 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 5641 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 5701 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5761 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 5821 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5881 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5941 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 6001 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 6061 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 6121 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 6181 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 6241 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 6301 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6361 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 6421 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 6481 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 6541 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6601 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6661 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6721 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6781 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6841 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 6901 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 6961 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 7021 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 7081 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 7141 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 7201 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7261 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7321 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7381 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7441 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7501 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7561 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7621 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7681 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7741 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7801 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7861 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7921 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7981 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 8041 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 8101 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 8161 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 8221 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8281 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8341 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8401 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8461 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8521 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8581 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8641 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8701 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8761 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8821 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8881 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8941 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 9001 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 9061 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 9121 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 9181 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9241 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9301 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9361 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9421 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9481 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9541 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9601 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9661 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9721 cagtgccaag ctg //