LOCUS V036225 9298 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036225 VERSION V036225 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..3330 /label="TFAP4(NM_003223)" /note="TFAP4(NM_003223)" /gene="TFAP4" CDS 3331..3396 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3440..3651 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3664..3932 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3961..3970 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3991..4653 /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 4723..4776 /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 4777..5376 /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 5377..5965 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6039..6272 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6344..6465 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6505..6640 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6673..6689) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6697..6713 /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(6721..6751) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6766..6787 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7075..7663) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7834..8694) /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(8695..8799) /gene="bla" /label="AmpR promoter" primer_bind 9273..9289 /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 agtatttcat ggtgcccact 2341 cagaaggtgc cctctttgca acatttcagg aaaacagaga aagaagtgat aggagggctc 2401 tgtagccttg ccaacattcc actaaccccc gagactcagc gggaccagga gcggcggatt 2461 cggcgggaga tcgccaacag caacgagcgg agacgcatgc agagcatcaa cgcgggattc 2521 cagtccctca agaccctcat cccccacaca gacggagaga agctcagcaa ggcagccatt 2581 ctccagcaga cagccgagta catcttctcc ctggagcagg agaagaccag gctcttgcag 2641 cagaacacac agctcaagcg cttcatccag gagctgagcg gctcgtcccc caagcgacgg 2701 cgggcagagg acaaggacga aggcataggc tccccggaca tctgggagga cgagaaggcg 2761 gaggacctgc ggcgggagat gattgagctg cggcagcagc tggacaagga gcgctcggtg 2821 cgcatgatgc tggaggagca ggtgcgctcg ctggaggccc acatgtaccc ggaaaagctc 2881 aaggtgattg cgcagcaggt gcagctgcag cagcagcagg aacaggtgag gctgctgcac 2941 caggagaagc tggagcggga acagcagcag ctgcggaccc agcttctgcc ccctccggcc 3001 cccacccacc accccacggt gatcgtgcca gcaccgcctc ctcctccctc ccaccacatc 3061 aatgtcgtca ccatgggccc ctcctcggtc atcaactctg tttccacatc ccggcaaaat 3121 ctggacacca tcgtgcaggc aatccagcac atcgagggca cccaggaaaa gcaggagctg 3181 gaggaggagc agcggcgagc tgtcatcgtg aagcctgtcc gcagctgccc ggaggccccc 3241 acctctgaca ccgcctccga ctccgaggcc tcagacagtg acgccatgga ccagagccgg 3301 gaggagccgt cgggggacgg ggagcttccc gactacaaag accatgacgg tgattataaa 3361 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3421 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3481 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3541 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3601 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3661 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3721 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3781 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3841 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3901 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 3961 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 4021 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 4081 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 4141 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4201 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4261 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4321 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4381 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4441 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4501 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4561 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4621 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4681 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4741 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 4801 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 4861 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 4921 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 4981 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 5041 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 5101 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 5161 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5221 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5281 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5341 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5401 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5461 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5521 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5581 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5641 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5701 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5761 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 5821 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 5881 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 5941 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 6001 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 6061 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 6121 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6181 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6241 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6301 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6361 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6421 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6481 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6541 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6601 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6661 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6721 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6781 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6841 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6901 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6961 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 7021 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 7081 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7141 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7201 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7261 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7321 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7381 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7441 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7501 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7561 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7621 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7681 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7741 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7801 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7861 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7921 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7981 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 8041 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 8101 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8161 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8221 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8281 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8341 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8401 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8461 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8521 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8581 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8641 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8701 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8761 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8821 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8881 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8941 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 9001 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 9061 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9121 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9181 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9241 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //