LOCUS V034972 9308 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034972 VERSION V034972 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 2305..2322 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" CDS 2329..4194 /label="F2(NM_000506)" /note="F2(NM_000506)" /gene="F2" CDS 4201..4227 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 4278..4489 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4502..4770 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4797..5396 /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 5403..5991 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6065..6298 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6370..6491 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6510..6645 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6683..6699) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6707..6723 /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(6731..6761) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6776..6797 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7085..7673) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7844..8704) /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(8705..8809) /gene="bla" /label="AmpR promoter" primer_bind 9283..9299 /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 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgcaccat catcatcatc atgatatcat ggcgcacgtc 2341 cgaggcttgc agctgcctgg ctgcctggcc ctggctgccc tgtgtagcct tgtgcacagc 2401 cagcatgtgt tcctggctcc tcagcaagca cggtcgctgc tccagcgggt ccggcgagcc 2461 aacaccttct tggaggaggt gcgcaagggc aacctggagc gagagtgcgt ggaggagacg 2521 tgcagctacg aggaggcctt cgaggctctg gagtcctcca cggctacgga tgtgttctgg 2581 gccaagtaca cagcttgtga gacagcgagg acgcctcgag ataagcttgc tgcatgtctg 2641 gaaggtaact gtgctgaggg tctgggtacg aactaccgag ggcatgtgaa catcacccgg 2701 tcaggcattg agtgccagct atggaggagt cgctacccac ataagcctga aatcaactcc 2761 actacccatc ctggggccga cctacaggag aatttctgcc gcaaccccga cagcagcacc 2821 acgggaccct ggtgctacac tacagacccc accgtgagga ggcaggaatg cagcatccct 2881 gtctgtggcc aggatcaagt cactgtagcg atgactccac gctccgaagg ctccagtgtg 2941 aatctgtcac ctccattgga gcagtgtgtc cctgatcggg ggcagcagta ccaggggcgc 3001 ctggcggtga ccacacatgg gctcccctgc ctggcctggg ccagcgcaca ggccaaggcc 3061 ctgagcaagc accaggactt caactcagct gtgcagctgg tggagaactt ctgccgcaac 3121 ccagacgggg atgaggaggg cgtgtggtgc tatgtggccg ggaagcctgg cgactttggg 3181 tactgcgacc tcaactattg tgaggaggcc gtggaggagg agacaggaga tgggctggat 3241 gaggactcag acagggccat cgaagggcgt accgccacca gtgagtacca gactttcttc 3301 aatccgagga cctttggctc gggagaggca gactgtgggc tgcgacctct gttcgagaag 3361 aagtcgctgg aggacaaaac cgaaagagag ctcctggaat cctacatcga cgggcgcatt 3421 gtggagggct cggatgcaga gatcggcatg tcaccttggc aggtgatgct tttccggaag 3481 agtccccagg agctgctgtg tggggccagc ctcatcagtg accgctgggt cctcaccgcc 3541 gcccactgcc tcctgtaccc gccctgggac aagaacttca ccgagaatga ccttctggtg 3601 cgcattggca agcactcccg caccaggtac gagcgaaaca ttgaaaagat atccatgttg 3661 gaaaagatct acatccaccc caggtacaac tggcgggaga acctggaccg ggacattgcc 3721 ctgatgaagc tgaagaagcc tgttgccttc agtgactaca ttcaccctgt gtgtctgccc 3781 gacagggaga cggcagccag cttgctccag gctggataca aggggcgggt gacaggctgg 3841 ggcaacctga aggagacgtg gacagccaac gttggtaagg ggcagcccag tgtcctgcag 3901 gtggtgaacc tgcccattgt ggagcggccg gtctgcaagg actccacccg gatccgcatc 3961 actgacaaca tgttctgtgc tggttacaag cctgatgaag ggaaacgagg ggatgcctgt 4021 gaaggtgaca gtgggggacc ctttgtcatg aagagcccct ttaacaaccg ctggtatcaa 4081 atgggcatcg tctcatgggg tgaaggctgt gaccgggatg ggaaatatgg cttctacaca 4141 catgtgttcc gcctgaagaa gtggatacag aaggtcattg atcagtttgg agagctcgag 4201 tacccatacg acgtaccaga ttacgcttag ggatccgcgg ccgcgaagga tctgcgatcg 4261 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg 4321 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga 4381 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt 4441 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg 4501 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg 4561 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta 4621 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc 4681 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt 4741 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga 4801 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca 4861 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc 4921 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 4981 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 5041 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 5101 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 5161 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 5221 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 5281 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 5341 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg 5401 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 5461 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 5521 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 5581 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 5641 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 5701 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 5761 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc 5821 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 5881 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 5941 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct 6001 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 6061 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg 6121 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6181 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6241 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 6301 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 6361 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 6421 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 6481 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 6541 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 6601 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 6661 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 6721 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 6781 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 6841 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 6901 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 6961 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 7021 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 7081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 7141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 7201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 7261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 7321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 7381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 7441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 7501 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 7561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 7621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 7681 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 7741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 7801 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 7861 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 7921 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 7981 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 8041 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 8101 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 8161 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8221 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 8281 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 8341 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 8401 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 8461 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 8521 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 8581 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 8641 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 8701 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 8761 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 8821 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 8881 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 8941 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 9001 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 9061 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 9121 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 9181 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 9241 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 9301 ccaagctg //