LOCUS V042805 9123 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042805 VERSION V042805 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" CDS 2297..3142 /label="ESD(NM_001984)" /note="ESD(NM_001984)" /gene="ESD" CDS 3149..3238 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" promoter 3282..3493 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3506..3774 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3803..3812 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3833..4495 /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 4565..4618 /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 4619..5218 /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 5219..5806 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5880..6113 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6185..6306 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6325..6460 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6498..6514) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6522..6538 /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(6546..6576) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6591..6612 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6900..7488) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7659..8519) /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(8520..8624) /gene="bla" /label="AmpR promoter" primer_bind 9098..9114 /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 tagcgaattc gccaccatgg cattgaagca gatttccagc aacaagtgct ttgggggatt 2341 gcagaaagtt tttgaacatg acagtgttga actaaactgc aaaatgaaat ttgctgtcta 2401 cttaccacca aaggcagaaa caggaaagtg ccctgcactg tattggctct caggtttaac 2461 ttgcacagag caaaatttta tatcaaaatc tggttatcat cagtctgctt cagaacatgg 2521 tcttgttgtc attgctccag ataccagccc tcgtggctgc aatattaaag gtgaagatga 2581 gagctgggac tttggcactg gtgctggatt ttatgttgat gccactgaag atccttggaa 2641 aaccaactac agaatgtact cttatgtcac agaggagctt ccccaactca taaatgccaa 2701 ttttccagtg gatccccaaa ggatgtctat ttttggccac tccatgggag gtcatggagc 2761 tctgatctgt gctttgaaaa atcctggaaa atacaaatct gtgtcagcat ttgctccaat 2821 ttgcaaccct gtactctgtc cctggggcaa aaaagccttt agtggatatt tgggaacaga 2881 tcaaagtaaa tggaaggctt atgatgctac ccaccttgtg aaatcctatc caggatctca 2941 gctggacata ctaattgatc aagggaaaga tgaccagttt cttttagatg gacagttact 3001 ccctgataac ttcatagctg cctgtacaga aaagaaaatc cccgttgttt ttcgattgca 3061 agagggttat gatcatagct actacttcat tgcaaccttt attactgacc acatcagaca 3121 tcatgctaaa tacctgaatg caggatccga acaaaaacta atatcagagg aagatttgga 3181 acagaaacta atcagtgaag aagatttaga gcaaaagtta atttccgagg aggacttata 3241 gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc 3301 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg 3361 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg 3421 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc 3481 cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc 3541 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc 3601 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg 3661 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct 3721 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct 3781 gtgaccggcg cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat 3841 ggagatcgag tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg 3901 cggagagggc acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc 3961 cctgaccttc agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg 4021 cacctacccc agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac 4081 caacacccgc atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg 4141 ctacgaggcc ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga 4201 cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc 4261 catgggcgat aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg 4321 ctactacagc ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat 4381 cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac 4441 cgagctgggc atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc 4501 ccgcgctcag tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc 4561 tcgcgagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat 4621 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg 4681 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg 4741 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 4801 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 4861 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 4921 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 4981 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 5041 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 5101 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 5161 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa 5221 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 5281 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 5341 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5401 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5461 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5521 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5581 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc 5641 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5701 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtctccg 5761 ccttcgccct cagacgagtc ggatctccct ttggccgcct ccccgcctgg tacctttaag 5821 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact 5881 ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta ctgggtctct 5941 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 6001 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 6061 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag 6121 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga 6181 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 6241 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 6301 atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc cgcccattct 6361 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6421 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagacggc 6481 ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 6541 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 6601 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 6661 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 6721 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 6781 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 6841 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 6901 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 6961 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 7021 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 7081 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 7141 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 7201 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 7261 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 7321 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 7381 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 7441 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 7501 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 7561 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 7621 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 7681 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 7741 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 7801 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 7861 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 7921 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 7981 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 8041 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 8101 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 8161 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 8221 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 8281 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 8341 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 8401 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 8461 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 8521 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 8581 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 8641 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt 8701 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg 8761 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt 8821 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag 8881 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga 8941 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg 9001 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta 9061 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag 9121 ctg //