LOCUS V022445 9502 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022445 VERSION V022445 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" regulatory 2820..2829 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2829..2852 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2853..3566 /label="Cd63(NM_007653)" /note="Cd63(NM_007653)" /gene="Cd63" CDS 3600..4310 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK*" /label="mCherry" promoter 4337..4836 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4857..5456 /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 5470..6058 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6265..6898 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7027..7043) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7051..7067 /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(7075..7105) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7120..7141 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7429..8017) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8188..9048) /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(9049..9153) /gene="bla" /label="AmpR promoter" polyA_signal 9201..9335 /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 ccaccatgga ttacaaggat gacgacgata agatggcggt ggaaggagga atgaagtgtg 2881 tcaagttttt gctctacgtt ctcctgctgg ccttctgcgc ctgtgcagtg ggattgatcg 2941 ccattggtgt agcggttcag gttgtcttga agcaggccat tacccatgag actactgctg 3001 gctcgctgtt gcctgtggtc atcattgcag tgggtgcctt cctcttcctg gtggcctttg 3061 tgggctgctg tggggcctgc aaggagaact actgtctcat gattacattt gccatcttcc 3121 tgtctcttat catgcttgtg gaggtggctg tggccattgc tggctatgtg tttagagacc 3181 aggtgaagtc agagtttaat aaaagcttcc agcagcagat gcagaattac cttaaagaca 3241 acaaaacagc cactattttg gacaaattgc agaaagaaaa taactgctgt ggagcttcta 3301 actacacaga ctgggaaaac atccccggca tggccaagga cagagtcccc gattcttgct 3361 gcatcaacat aactgtgggc tgtgggaatg atttcaagga atccactatc catacccagg 3421 gctgcgtgga gactatagca atatggctaa ggaagaacat actgctggtg gctgcagcgg 3481 ccctgggcat tgcttttgtg gaggtcttgg gaattatctt ctcctgctgt ctggtgaaga 3541 gtattcgaag tggctatgaa gtaatgggag gaggtggatc ggatccaccg gtcgccacca 3601 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg 3661 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc 3721 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct 3781 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc 3841 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg 3901 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg 3961 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa 4021 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg 4081 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg 4141 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca 4201 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac 4261 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtag cggccgcgac 4321 tctagataat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct 4381 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc 4441 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct 4501 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga 4561 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg 4621 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct 4681 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg 4741 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc 4801 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag cttaccatga 4861 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca 4921 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc 4981 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 5041 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 5101 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 5161 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 5221 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 5281 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 5341 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 5401 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaccgc 5461 gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 5521 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5581 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 5641 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5701 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5761 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5821 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 5881 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5941 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 6001 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 6061 ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag 6121 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc 6181 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc 6241 actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata 6301 tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag aactacacac 6361 cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg 6421 agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga 6481 gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc 6541 tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc 6601 gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga 6661 ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg 6721 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6781 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6841 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 6901 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 6961 agtgagaggc cttgacattg ctagcgtttt accgtcgacc tctagctaga gcttggcgta 7021 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 7081 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 7141 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 7201 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 7261 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 7321 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 7381 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 7441 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 7501 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 7561 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 7621 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 7681 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 7741 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 7801 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 7861 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 7921 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 7981 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 8041 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 8101 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 8161 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 8221 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 8281 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 8341 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 8401 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 8461 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 8521 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 8581 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 8641 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 8701 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 8761 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 8821 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 8881 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 8941 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 9001 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 9061 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 9121 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 9181 cgtcgacgga tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca 9241 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 9301 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa 9361 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt 9421 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa 9481 tatgtactac aaacttagta gt //