LOCUS V044494 9478 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044494 VERSION V044494 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..3504 /label="APOA4(NM_000482)" /note="APOA4(NM_000482)" /gene="APOA4" CDS 3511..3576 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3620..3831 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3844..4112 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4141..4150 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4171..4833 /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 4903..4956 /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 4957..5556 /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 5557..6145 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6219..6452 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6524..6645 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6685..6820 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6853..6869) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6877..6893 /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(6901..6931) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6946..6967 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7255..7843) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8014..8874) /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(8875..8979) /gene="bla" /label="AmpR promoter" primer_bind 9453..9469 /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 gccaccatgt tcctgaaggc cgtggtcctg 2341 accctggccc tggtggctgt cgccggagcc agggctgagg tcagtgctga ccaggtggcc 2401 acggtgatgt gggactactt cagccagctg agcaacaatg ccaaggaggc cgtggaacat 2461 ctccagaaat ctgaactcac ccagcaactc aatgccctct tccaggacaa acttggagaa 2521 gtgaacactt acgcaggtga cctgcagaag aagctggtgc cctttgccac cgagctgcat 2581 gaacgcctgg ccaaggactc ggagaaactg aaggaggaga ttgggaagga gctggaggag 2641 ctgagggccc ggctgctgcc ccatgccaat gaggtgagcc agaagatcgg ggacaacctg 2701 cgagagcttc agcagcgcct ggagccctac gcggaccagc tgcgcaccca ggtcagcacg 2761 caggccgagc agctgcggcg ccagctgacc ccctacgcac agcgcatgga gagagtgctg 2821 cgggagaacg ccgacagcct gcaggcctcg ctgaggcccc acgccgacga gctcaaggcc 2881 aagatcgacc agaacgtgga ggagctcaag ggacgcctta cgccctacgc tgacgaattc 2941 aaagtcaaga ttgaccagac cgtggaggag ctgcgccgca gcctggctcc ctatgctcag 3001 gacacgcagg agaagctcaa ccaccagctt gagggcctga ccttccagat gaagaagaac 3061 gccgaggagc tcaaggccag gatctcggcc agtgccgagg agctgcggca gaggctggcg 3121 cccttggccg aggacgtgcg tggcaacctg aggggcaaca ccgaggggct gcagaagtca 3181 ctggcagagc tgggtgggca cctggaccag caggtggagg agttccgacg ccgggtggag 3241 ccctacgggg aaaacttcaa caaagccctg gtgcagcaga tggaacagct caggcagaaa 3301 ctgggccccc atgcggggga cgtggaaggc cacttgagct tcctggagaa ggacctgagg 3361 gacaaggtca actccttctt cagcaccttc aaggagaaag agagccagga caagactctc 3421 tccctccctg agctggagca acagcaggaa cagcagcagg agcagcagca ggagcaggtg 3481 cagatgctgg cccctttgga gagcggatcc gactacaaag accatgacgg tgattataaa 3541 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3601 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3661 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3721 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3781 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3841 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3901 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3961 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4021 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4081 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 4141 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 4201 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 4261 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 4321 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4381 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4441 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4501 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4561 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4621 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4681 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4741 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4801 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4861 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4921 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 4981 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 5041 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 5101 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 5161 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 5221 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 5281 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 5341 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5401 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5461 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5521 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5581 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5641 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5701 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5761 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5821 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5881 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5941 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 6001 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 6061 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 6121 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 6181 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 6241 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 6301 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6361 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6421 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6481 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6541 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6601 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6661 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6721 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6781 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6841 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6901 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6961 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 7021 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 7081 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 7141 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 7201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 7261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7681 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7801 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7861 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7921 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7981 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 8041 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 8101 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 8161 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 8221 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 8281 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8341 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8401 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8461 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8521 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8581 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8641 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8701 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8761 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8821 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8881 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8941 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 9001 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 9061 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 9121 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 9181 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 9241 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9301 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9361 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9421 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //