LOCUS V021753 8950 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021753 VERSION V021753 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2301..3875 /label="HRG(NM_000412)" /note="HRG(NM_000412)" /gene="HRG" promoter 3920..4131 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4144..4412 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4439..5038 /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 5045..5633 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5707..5940 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6012..6133 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6152..6287 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6325..6341) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6349..6365 /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(6373..6403) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6418..6439 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6727..7315) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7486..8346) /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(8347..8451) /gene="bla" /label="AmpR promoter" primer_bind 8925..8941 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta 2281 gagctagcga attcgccacc atgaaggcac tcattgcagc actgcttttg atcacattgc 2341 agtattcgtg tgccgtgagt cccactgact gcagtgctgt tgagccggag gctgagaaag 2401 ctctagacct gatcaataaa aggcgacggg atggctacct tttccaattg ctgcggattg 2461 ctgatgccca cttggacaga gtggaaaata caactgtata ttacttagtc ttagatgtgc 2521 aagaatcgga ctgttcggtc ctatccagga aatactggaa tgactgtgag ccacctgatt 2581 ccagacgtcc atctgaaata gtgatcggac aatgtaaggt aatagctaca agacattccc 2641 atgaatctca ggacctcaga gtgattgact ttaactgcac cacaagttct gtctcttcag 2701 cactggccaa taccaaagat agtccggtcc tcatagattt ctttgaggat actgagcgct 2761 acagaaaaca agccaacaaa gcccttgaga agtacaaaga ggagaatgat gactttgcct 2821 ctttcagagt ggaccgaatc gagagagttg caagagtgag aggaggggaa ggaactggtt 2881 acttcgtgga cttctctgtg cggaactgcc ccagacacca tttccccaga caccccaatg 2941 tctttggatt ctgcagagca gatttgttct atgatgtaga agccttggac ttggaaagcc 3001 cgaaaaacct tgtcataaac tgtgaagtct tcgaccctca ggaacatgag aacatcaatg 3061 gtgtaccgcc tcatttggga catcccttcc actggggtgg gcatgagcgt tcttctacca 3121 ccaagcctcc attcaagccc catggatcta gagatcatca tcatccccac aagccacacg 3181 aacatggacc cccacctcct ccagatgaaa gagatcactc acatggaccc ccacttccac 3241 aaggccctcc tccactattg cccatgtcct gctcaagttg tcaacatgcc acttttggca 3301 caaatggggc ccaaagacat tctcataata ataattccag tgacctccat ccccataagc 3361 atcattccca tgaacagcat ccccacggac accatcccca tgcacaccat cctcatgaac 3421 atgataccca tagacagcat ccccatggac accaccccca tggacaccat cctcatggac 3481 accaccccca tggacaccat ccccatggac accatcccca ctgccatgat ttccaagact 3541 atggaccttg tgacccacca ccccataacc aaggtcactg ttgccatggc cacggcccac 3601 cacctgggca cttaagaagg cgaggcccag gtaaaggacc ccgtcccttc cattgcagac 3661 aaattggatc tgtgtaccga ctccctcctc taagaaaagg tgaggtgctg ccacttcctg 3721 aggccaattt tcccagcttc ccattgccgc accacaaaca tcctctaaag ccagacaatc 3781 agccctttcc tcaatcagtc tctgaatcat gtccagggaa gttcaagagt gggtttccac 3841 aagtttccat gttttttaca catacatttc caaaataagc ggccgcgaag gatctgcgat 3901 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3961 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 4021 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 4081 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 4141 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 4201 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4261 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4321 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4381 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat 4441 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg 4501 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg 4561 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 4621 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 4681 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 4741 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 4801 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 4861 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 4921 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 4981 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt 5041 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 5101 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 5161 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5221 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5281 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5341 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5401 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 5461 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5521 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5581 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 5641 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 5701 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 5761 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 5821 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 5881 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 5941 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 6001 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 6061 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 6121 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 6181 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 6241 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 6301 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6361 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6421 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6481 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6541 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6601 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6661 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6721 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6781 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6841 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6901 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6961 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7021 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7081 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7141 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7201 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7261 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7321 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7381 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7441 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7501 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7561 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7621 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7681 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7741 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7801 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7861 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7921 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7981 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8041 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8101 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8161 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8221 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8281 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8341 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8401 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8461 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8521 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8581 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8641 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8701 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8761 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8821 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8881 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 8941 tgccaagctg //