LOCUS V024028 10004 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024028 VERSION V024028 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..4047 /label="IGF2BP1(NM_006546)" /note="IGF2BP1(NM_006546)" /gene="IGF2BP1" CDS 4054..4119 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4163..4374 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4387..4655 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4684..4693 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4714..5376 /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 5446..5499 /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 5500..6099 /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 6100..6687 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6761..6994 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7066..7187 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7206..7341 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7379..7395) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7403..7419 /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(7427..7457) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7472..7493 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7781..8369) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8540..9400) /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(9401..9505) /gene="bla" /label="AmpR promoter" primer_bind 9979..9995 /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 gccaccatga acaagcttta catcggcaac 2341 ctcaacgaga gcgtgacccc cgcggacttg gagaaagtgt ttgcggagca caagatctcc 2401 tacagcggcc agttcttggt caaatccggc tacgccttcg tggactgccc ggacgagcac 2461 tgggcgatga aggccatcga aactttctcc gggaaagtag aattacaagg aaaacgctta 2521 gagattgaac attcggtgcc caaaaaacaa aggagccgga aaattcaaat ccgaaatatt 2581 ccaccccagc tccgatggga agtactggac agcctgctgg ctcagtatgg tacagtagag 2641 aactgtgagc aagtgaacac cgagagtgag acggcagtgg tgaatgtcac ctattccaac 2701 cgggagcaga ccaggcaagc catcatgaag ctgaatggcc accagttgga gaaccatgcc 2761 ctgaaggtct cctacatccc cgatgagcag atagcacagg gacctgagaa tgggcgccga 2821 gggggctttg gctctcgggg tcagccccgc cagggctcac ctgtggcagc gggggcccca 2881 gccaagcagc agcaagtgga catccccctt cggctcctgg tgcccaccca gtatgtgggt 2941 gccattattg gcaaggaggg ggccaccatc cgcaacatca caaaacagac ccagtccaag 3001 atagacgtgc ataggaagga gaacgcaggt gcagctgaaa aagccatcag tgtgcactcc 3061 acccctgagg gctgctcctc cgcttgtaag atgatcttgg agattatgca taaagaggct 3121 aaggacacca aaacggctga cgaggttccc ctgaagatcc tggcccataa taactttgta 3181 gggcgtctca ttggcaagga aggacggaac ctgaagaagg tagagcaaga taccgagaca 3241 aaaatcacca tctcctcgtt gcaagacctt accctttaca accctgagag gaccatcact 3301 gtgaaggggg ccatcgagaa ttgttgcagg gccgagcagg aaataatgaa gaaagttcgg 3361 gaggcctatg agaatgatgt ggctgccatg agcctgcagt ctcacctgat ccctggcctg 3421 aacctggctg ctgtaggtct tttcccagct tcatccagcg cagtcccgcc gcctcccagc 3481 agcgttactg gggctgctcc ctatagctcc tttatgcagg ctcccgagca ggagatggtg 3541 caggtgttta tccccgccca ggcagtgggc gccatcatcg gcaagaaggg gcagcacatc 3601 aaacagctct cccggtttgc cagcgcctcc atcaagattg caccacccga aacacctgac 3661 tccaaagttc gtatggttat catcactgga ccgccagagg cccaattcaa ggctcaggga 3721 agaatctatg gcaaactcaa ggaggagaac ttctttggtc ccaaggagga agtgaagctg 3781 gagacccaca tacgtgtgcc agcatcagca gctggccggg tcattggcaa aggtggaaaa 3841 acggtgaacg agttgcagaa tttgacggca gctgaggtgg tagtaccaag agaccagacc 3901 cctgatgaga acgaccaggt catcgtgaaa atcatcggac atttctatgc cagtcagatg 3961 gctcaacgga agatccgaga catcctggcc caggttaagc agcagcatca gaagggacag 4021 agtaaccagg cccaggcacg gaggaaggga tccgactaca aagaccatga cggtgattat 4081 aaagatcatg acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc 4141 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 4201 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 4261 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 4321 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 4381 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 4441 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 4501 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 4561 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 4621 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 4681 gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc 4741 accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag 4801 cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac 4861 ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac 4921 gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag 4981 tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg 5041 atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac 5101 aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg 5161 gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg 5221 gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc 5281 atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag 5341 taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat 5401 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt 5461 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg 5521 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 5581 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 5641 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 5701 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 5761 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 5821 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 5881 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 5941 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 6001 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 6061 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa 6121 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 6181 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 6241 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 6301 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 6361 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 6421 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 6481 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 6541 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 6601 cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt 6661 cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 6721 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 6781 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 6841 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 6901 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 6961 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 7021 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 7081 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 7141 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 7201 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 7261 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 7321 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 7381 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 7441 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 7501 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 7561 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 7621 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7681 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7741 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7801 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 8041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 8101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 8161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 8221 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 8281 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 8341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 8401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 8461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 8521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 8581 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 8641 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 8701 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 8761 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8821 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8881 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8941 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 9001 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 9061 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 9121 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 9181 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 9241 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 9301 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 9361 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 9421 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 9481 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 9541 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 9601 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 9661 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 9721 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 9781 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 9841 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 9901 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 9961 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //