LOCUS V029553 9178 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029553 VERSION V029553 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..3204 /label="Echdc2(NM_026728)" /note="Echdc2(NM_026728)" /gene="Echdc2" CDS 3211..3276 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3320..3531 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3544..3812 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3841..3850 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3871..4533 /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 4603..4656 /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 4657..5256 /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 5257..5845 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5919..6152 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6224..6345 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6385..6520 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6553..6569) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6577..6593 /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(6601..6631) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6646..6667 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6955..7543) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7714..8574) /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(8575..8679) /gene="bla" /label="AmpR promoter" primer_bind 9153..9169 /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 gccaccatgc tacgcgtact gccccgcgcg 2341 ctgcgcctcc cgtgctcctg gaggttctcg ggggcccggg actgcgcctc tcacgcgacg 2401 accaggacac ccgaaatcca agtgcaggct ctgacaggtc ccaaccaagg aatcactgag 2461 attctgatga acagacctaa tgcccgcaat gccttgggga atgtgtttgt tagtgagctg 2521 ctggaagctc tggcccagct tcgggaagac cagcaagtcc gggtcctgct cttcagaagt 2581 gcggtgaagg gagtgttctg tgcaggtgca gacctgaagg agcgggagca gatgagtgat 2641 gtggaggtgg ggacctttgt ccagcggctc cgaggcctga tgagtgagat cgcagccttc 2701 cctgtgccca ccattgcagc catggatggg tttgccttag gcgggggctt ggaacttgcc 2761 ctggcctgtg acctccgaat agcagcttct tcggctgtca tggggctaat tgagaccact 2821 cgaggacttc tcccaggagc aggagggact cagaggctgc ctcgctgcct gggcgtggcc 2881 ctagcgaagg agctcatctt cacaggccgg cgactgaacg gggcacaggc ccgtgagttg 2941 ggcctggtaa atcatgccgt ggctcagaat gaggagggta acgctgccta ccaccgggcc 3001 ctggcactgg cacaggagat cctgccccag gctcccattg ctgtgcgcct gggcaaagtt 3061 gccatcgaca gaggaatgga ggtggacatt gcatcaggga tggccataga acagatgtgc 3121 tatgcccaga acatcccaac ccaggacagg ctggaaggaa tggcagcctt cagggagaaa 3181 cgagctccca aatttgtggg caagggatcc gactacaaag accatgacgg tgattataaa 3241 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3301 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3361 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3421 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3481 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3541 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3601 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3661 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3721 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3781 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 3841 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 3901 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 3961 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 4021 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4081 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4141 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4201 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4261 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4321 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4381 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4441 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4501 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4561 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4621 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 4681 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 4741 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 4801 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 4861 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 4921 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 4981 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 5041 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5101 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5161 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5221 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5281 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5341 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5401 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5461 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5521 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5581 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5641 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 5701 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 5761 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 5821 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 5881 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 5941 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 6001 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6061 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6121 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6181 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6241 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6301 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6361 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6421 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6481 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6541 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6601 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6661 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6721 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6781 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6841 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6901 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6961 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7021 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7081 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7141 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7201 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7261 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7321 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7381 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7441 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7501 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7561 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7621 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7681 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7741 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7801 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7861 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 7921 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7981 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8041 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8101 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8161 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8221 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8281 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8341 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8401 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8461 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8521 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8581 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8641 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8701 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8761 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8821 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 8881 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 8941 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9001 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9061 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9121 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //