LOCUS V039334 10312 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039334 VERSION V039334 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" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2843..4999 /label="HOOK2(NM_013312)" /note="HOOK2(NM_013312)" /gene="HOOK2" CDS 5015..5080 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 5100..5599 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5620..6219 /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 6233..6821 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7028..7661 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7790..7806) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7814..7830 /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(7838..7868) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7883..7904 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8192..8780) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8951..9811) /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(9812..9916) /gene="bla" /label="AmpR promoter" polyA_signal 9964..10098 /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 cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 ctcaagcttc gaattcgcca ccatgagcgt ggacaaagct gagctatgcg ggtctctgct 2881 cacctggtta cagacgttcc acgttccgtc tccctgtgcc agccctcagg acctgagcag 2941 cggccttgcc gtagcctatg tgctgaacca gatagacccc tcctggttca acgaggcatg 3001 gctccagggc atctcggaag atccaggtcc caactggaag ctgaaggtca gcaatctgaa 3061 gatggtctta cggagcctag tagagtactc ccaggatgtc ctggcgcatc ctgtgtcaga 3121 agagcatctc ccagatgtga gcctcattgg agagttctca gacccggcag agctcggcaa 3181 gctgcttcag ctggtgctgg gctgtgccat cagttgcgag aaaaagcagg accacatcca 3241 gagaatcatg acgctggaag aatcggttca gcatgtggtg atggaagcca tccaagagct 3301 catgaccaaa gacactcctg actccctgtc accagagacg tatggcaact ttgacagcca 3361 gtcccgcagg tactatttcc taagtgagga ggctgaggag ggggacgaat tacagcagcg 3421 ctgtctggat ctggagcggc agctgatgct cctgtcagag gagaagcaga gcctggcgca 3481 agagaatgca gggctgcggg agcggatggg ccggcctgaa ggcgagggta ccccaggtct 3541 cactgccaag aagctgctgc tgctgcaatc ccagctggag cagttgcagg aggagaactt 3601 caggctggag agtggcaggg aggatgagcg cctgcgctgt gccgagctgg agagggaggt 3661 tgcggagctg cagcaccgga accaggcgct gactagcctg gcccaggagg cacaggccct 3721 gaaggatgag atggatgaac tacggcagtc ttcggagcgt gctgggcagc tggaggccac 3781 gctgaccagt tgccggcgcc gcttgggcga gctgagggag ctgcggcggc aggtgcggca 3841 gctggaggaa cgcaacgccg gccacgccga gcgcacgcga caactggagg atgagctacg 3901 ccgagcgggc tccctgcgcg cccagctgga ggcgcagcgg cggcaggtgc aggaactgca 3961 gggccagcgg caggaggagg ccatgaaggc cgagaaatgg ctatttgaat gccgcaacct 4021 ggaggaaaag tatgagtcgg tgacaaagga gaaggagcgg ctgttggcgg agcgggactc 4081 cttgcgggag gccaatgagg agctgcgctg cgcccagctg cagccgcggg ggttgaccca 4141 ggccgatccc tcactggatc ccacctccac acccgtggat aacttagccg cagagatcct 4201 gcctgcggag ctcagggaga cgctcctgcg gcttcagctg gagaacaagc ggctgtgcag 4261 gcaggaggcg gccgaccggg agcggcagga ggagctgcag cgccacctgg aggatgccaa 4321 ccgcgcgcgc cacgggttgg agacgcagca ccggctgaac cagcagcagc tatccgagct 4381 gcgggcccag gtggaggacc tgcagaaagc cctgcaggag caggggggca agactgaaga 4441 tgccatttcc attttgctga aaaggaagct ggaggaacat ttgcagaagc ttcatgaggc 4501 agatctggag ttgcagagga agcgggagta cattgaggag ctggagccac ccactgacag 4561 cagcacagcc cggcggatcg aggagctgca gcataacttg cagaagaagg acgcggactt 4621 gcgggccatg gaggagcgat accgccgcta cgtggacaag gcccgcatgg tcatgcagac 4681 catggaaccc aagcagcggc cagctgcggg ggcacctcca gaactccatt ccctgaggac 4741 acagctccga gaacgggatg tccgcatccg acacctggag atggactttg agaaaagccg 4801 aagtcagcgg gagcaggaag aaaagctgct catcagtgcc tggtataata tgggcatggc 4861 cttgcagcag cgagctgggg aggagcgggc gcctgcccat gcccagtcat tcctggcaca 4921 gcagcggctg gcaaccaatt ctcgccgtgg acccttggga cgcctggcat ctctgaacct 4981 tcgccccact gacaagcacg gtggaggcgg atccgactac aaagaccatg acggtgatta 5041 taaagatcat gacatcgact acaaggatga cgatgacaag tagtctagat aattctaccg 5101 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg 5161 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg 5221 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag 5281 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg 5341 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg 5401 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg 5461 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc 5521 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc 5581 ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg 5641 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 5701 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 5761 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 5821 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 5881 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 5941 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 6001 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 6061 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 6121 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 6181 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac cgcgtctgga acaatcaacc 6241 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 6301 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 6361 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 6421 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 6481 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 6541 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 6601 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt 6661 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 6721 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 6781 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt 6841 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat 6901 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta 6961 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 7021 aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc 7081 taccacacac aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga 7141 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa 7201 gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat 7261 gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg 7321 gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga 7381 ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc 7441 ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca 7501 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 7561 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 7621 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 7681 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca 7741 ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 7801 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 7861 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 7921 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 7981 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 8041 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 8101 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 8161 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 8221 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 8281 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 8341 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 8401 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 8461 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 8521 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 8581 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 8641 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 8701 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 8761 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 8821 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 8881 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 8941 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 9001 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 9061 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 9121 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 9181 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 9241 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 9301 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 9361 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 9421 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 9481 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 9541 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 9601 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 9661 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 9721 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 9781 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 9841 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 9901 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 9961 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 10021 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 10081 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc 10141 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat 10201 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta 10261 gtagttttta aagaaattgt atttgttaaa tatgtactac aaacttagta gt //