LOCUS V025688 9004 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025688 VERSION V025688 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 2826..3671 /label="TMIGD2(NM_144615)" /note="TMIGD2(NM_144615)" /gene="TMIGD2" CDS 3672..3701 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 3702..3719 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" promoter 3792..4291 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4312..4911 /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 4925..5513 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5720..6353 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6482..6498) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6506..6522 /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(6530..6560) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6575..6596 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6884..7472) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7643..8503) /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(8504..8608) /gene="bla" /label="AmpR promoter" polyA_signal 8656..8790 /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 ccaccatggg gtccccgggc atggtgctgg gcctcctggt gcagatctgg gccctgcaag 2881 aagcctcaag cctgagcgtg cagcaggggc ccaacttgct gcaggtgagg cagggcagtc 2941 aggcgaccct ggtctgccag gtggaccagg ccacagcctg ggaacggctc cgtgttaagt 3001 ggacaaagga tggggccatc ctgtgtcaac cgtacatcac caacggcagc ctcagcctgg 3061 gggtctgcgg gccccaggga cggctctcct ggcaggcacc cagccatctc accctgcagc 3121 tggaccctgt gagcctcaac cacagcgggg cgtacgtgtg ctgggcggcc gtagagattc 3181 ctgagttgga ggaggctgag ggcaacataa caaggctctt tgtggaccca gatgacccca 3241 cacagaacag aaaccggatc gcaagcttcc caggattcct cttcgtgctg ctgggggtgg 3301 gaagcatggg tgtggctgcg atcgtgtggg gtgcctggtt ctggggccgc cgcagctgcc 3361 agcaaaggga ctcaggtaac agcccaggaa atgcattcta cagcaacgtc ctataccggc 3421 cccggggggc cccaaagaag agtgaggact gctctggaga ggggaaggac cagaggggcc 3481 agagcattta ttcaacctcc ttcccgcaac cggccccccg ccagccgcac ctggcgtcaa 3541 gaccctgccc cagcccgaga ccctgcccca gccccaggcc cggccacccc gtctctatgg 3601 tcagggtctc tcctagacca agccccaccc agcagccgag gccaaaaggg ttccccaaag 3661 tgggagagga ggagcagaag ctgatctcag aggaggacct gcaccatcac catcaccatt 3721 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatccc gcgactctag 3781 ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc 3841 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc 3901 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc 3961 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat 4021 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg 4081 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag 4141 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc 4201 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct 4261 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag 4321 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4381 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4441 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4501 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4561 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 4621 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 4681 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 4741 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 4801 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 4861 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg 4921 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4981 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 5041 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5101 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5161 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5221 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5281 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct 5341 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5401 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5461 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat 5521 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct 5581 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc 5641 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 5701 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt 5761 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg 5821 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca 5881 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg 5941 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca 6001 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct 6061 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg 6121 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct 6181 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 6241 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 6301 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag 6361 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga 6421 gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg gcgtaatcat 6481 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 6541 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 6601 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 6661 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 6721 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 6781 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 6841 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 6901 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 6961 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 7021 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 7081 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 7141 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 7201 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 7261 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 7321 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 7381 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 7441 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 7501 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 7561 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 7621 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 7681 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 7741 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 7801 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 7861 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 7921 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 7981 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 8041 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 8101 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 8161 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 8221 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 8281 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 8341 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 8401 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 8461 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 8521 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 8581 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 8641 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc 8701 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 8761 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat 8821 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc 8881 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt 8941 actacaaact tagtagtttt taaagaaatt gtatttgtta aatatgtact acaaacttag 9001 tagt //