LOCUS V042069 10159 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042069 VERSION V042069 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" promoter 2337..3519 /note="strong constitutive promoter for human elongation factor EF-1α" /label="EF-1α promoter" CDS 3548..4825 /label="STK25(NM_001271977)" /note="STK25(NM_001271977)" /gene="STK25" CDS 4832..4897 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4909..5495 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5515..6114 /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 6128..6716 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6923..7556 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7684..7700) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7708..7724 /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(7732..7762) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7777..7798 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8086..8674) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8845..9705) /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(9706..9810) /gene="bla" /label="AmpR promoter" polyA_signal 9858..9992 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg 2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc 2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct 2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtacgaatt 3541 cgccaccatg gctcacctcc ggggatttgc caaccagcac tctcgagtgg accctgagga 3601 gctcttcacc aagctcgacc gcattggcaa gggctcgttt ggggaggtct acaagggcat 3661 cgataaccac acaaaggagg tggtggccat caagatcatc gacctggagg aggccgagga 3721 tgagatcgag gacatccagc aggagatcac tgtcctcagt cagtgcgaca gcccctacat 3781 cacccgctac tttggctcct acctaaagag caccaagcta tggatcatca tggagtacct 3841 gggcggcggc tcagcactgg acttgcttaa accaggtccc ctggaggaga catacattgc 3901 cacgatcctg cgggagattc tgaagggcct ggattatctg cactccgaac gcaagatcca 3961 ccgagacatc aaagctgcca acgtgctact ctcggagcag ggtgacgtga agctggcgga 4021 ctttggggta gcagggcagc tcacagacac gcagattaag aggaacacat tcgtgggcac 4081 ccccttctgg atggcacctg aggtcatcaa gcagtcggcc tacgacttca aggctgacat 4141 ctggtccctg gggatcacag ccatcgagct ggccaagggg gagcctccaa actctgacct 4201 ccaccccatg cgcgtcctgt tcctgattcc caagaacagc ccacccacac tggagggcca 4261 gcacagcaag cccttcaagg agttcgtgga ggcctgcctc aacaaagacc cccgattccg 4321 gcccacggcc aaggagctcc tgaagcacaa gttcatcaca cgctacacca agaagacctc 4381 cttcctcacg gagctcatcg accgctataa gcgctggaag tcagaggggc atggcgagga 4441 gtccagctct gaggactctg acattgatgg cgaggcggag gacggggagc agggccccat 4501 ctggacgttc ccccctacca tccggccgag tccacacagc aagcttcaca aggggacggc 4561 cctgcacagt tcacagaagc ctgcggagcc cgtcaagagg cagccgaggt cccagtgcct 4621 gtccacgctg gtccggcccg tcttcggaga gctcaaagag aagcacaagc agagcggcgg 4681 gagcgtgggt gcgctggagg agctggagaa cgccttcagc ctggccgagg agtcctgccc 4741 cggcatctca gacaagctga tggtgcacct ggtggagcga gtgcagaggt tttcacacaa 4801 cagaaaccac ctgacatcca cccgcggccg cgactacaaa gaccatgacg gtgattataa 4861 agatcatgac atcgactaca aggatgacga tgacaagtag tctagacgcc cctctccctc 4921 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta 4981 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc 5041 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct 5101 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt 5161 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa 5221 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg 5281 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga 5341 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac 5401 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt 5461 cctttgaaaa acacgatgat aagcttgcca caacccacaa ggagacgacc ttccatgacc 5521 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc 5581 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac 5641 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 5701 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 5761 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5821 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5881 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5941 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 6001 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 6061 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctagacgcgt 6121 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 6181 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 6241 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 6301 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 6361 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 6421 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 6481 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg 6541 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 6601 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 6661 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 6721 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca 6781 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca 6841 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 6901 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc 6961 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca 7021 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag 7081 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc 7141 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta 7201 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga 7261 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact 7321 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc 7381 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 7441 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 7501 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 7561 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 7621 tgagaggcct tgacattgct agcgtttacc gtcgacctct agctagagct tggcgtaatc 7681 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 7741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 7801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 7861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 7921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 7981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 8041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 8101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 8161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 8221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 8281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 8341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 8401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 8461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 8521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 8581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 8641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 8701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 8761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 8821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 8881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 8941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 9001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 9061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 9121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 9181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 9241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 9301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 9361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 9421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 9481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 9541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 9601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 9661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 9721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 9781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 9841 cgacggatcg ggagatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata 9901 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 9961 aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc 10021 atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac 10081 tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat 10141 gtactacaaa cttagtagt //