LOCUS V032022 9667 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032022 VERSION V032022 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..4363 /label="GTDC1(NM_001376306)" /note="GTDC1(NM_001376306)" /gene="GTDC1" CDS 4370..4435 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4455..4954 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4975..5574 /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 5588..6176 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6383..7016 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7145..7161) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7169..7185 /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(7193..7223) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7238..7259 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7547..8135) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8306..9166) /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(9167..9271) /gene="bla" /label="AmpR promoter" polyA_signal 9319..9453 /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 ccatgagtat cctcatcatt gaagcattct atggaggctc 2881 ccataaacag ctggtggatc ttcttcaaga agagttagga gactgtgtcg tttataccct 2941 tcctgcaaag aaatggcatt ggagagcccg gacatctgct ttatatttct ctcagaccat 3001 tcccatcagt gagcattaca ggaccctctt tgcaagttca gtgcttaacc tgaccgaact 3061 ggctgccctt cggcctgacc ttgggaaact gaaaaagatt ctgtattttc acgagaacca 3121 gttgatatat cctgtcaaga aatgtcagga gagggatttc caatatggat acaaccaaat 3181 tctttcatgc ctggtggctg atgtggttgt attcaactca gtttttaata tggaatcatt 3241 tctcacttcc atgggaaaat ttatgaagct gattcctgat cacagaccca aggatctgga 3301 aagcatcatc agacccaagt gccaagttat ttactttccc atcaggtttc ctgatgtgag 3361 cagcacctcc accagagttg gaactgggaa agagaaagga agtgttcaga catgtgccta 3421 tttgcatagt cacgtcattg aattaatgcc aggacagcac aaacattctg tccagttcta 3481 tagaaaagcc agtttcaccc catcagtaag attcatgccc aagcacaaaa caacccattt 3541 aaagaagatg ctcggcctta aaggaaatgg cggtgcggtt ctgtccatgg cccttccttt 3601 tcagccagag cagagagatt cagaggattt attgaagaat tttaattcag agtgtgatac 3661 acactgtggc cttgatactg cacgacaaga atatttgggt aactcattaa gacaggaatc 3721 agacttgaaa aaatccacct cgtcagataa ttcaagctct catcatggtg aaaataaaca 3781 aaatctgact gttgatccct gtgacatttt gggtggagtt gataatcagc aaagactgct 3841 acacattgtc tggcctcaca ggtgggagca tgataaagat ccagaaagct tttttaaggt 3901 attaatgcat cttaaagact taggactcaa tttccacgtg tctgtacttg gagaaacctt 3961 cacagatgtc ccagatattt tttcagaggc caaaaaggca ttgggatctt ctgtcttaca 4021 ctggggctac ttacccagca aagatgacta tttccaagta ctgtgcatgg ctgatgttgt 4081 catctcaaca gctaagcatg aattctttgg agtggcaatg ttggaagctg tgtactgtgg 4141 gtgttaccca ctttgtccta aagatttggt ttatcccgaa atatttccag ctgaatatct 4201 gtattctaca cctgaacagc tttcaaaaag gctccagaat ttctgcaaga gaccagatat 4261 tataagaaaa catctctata agggtgaaat cgctccgttt tcttgggcag ccctacatgg 4321 taaattcagg tctctgctta caacagaacc tagggaagat ttgggatccg actacaaaga 4381 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtc 4441 tagataattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt 4501 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac 4561 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac 4621 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca 4681 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa 4741 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca 4801 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg 4861 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc 4921 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc 4981 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 5041 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 5101 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 5161 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 5221 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5281 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5341 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5401 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5461 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5521 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaccgcgt 5581 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5641 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5701 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5761 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 5821 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 5881 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 5941 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg 6001 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 6061 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 6121 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 6181 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca 6241 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca 6301 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 6361 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc 6421 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca 6481 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag 6541 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc 6601 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta 6661 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga 6721 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact 6781 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc 6841 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 6901 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 6961 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 7021 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 7081 tgagaggcct tgacattgct agcgttttac cgtcgacctc tagctagagc ttggcgtaat 7141 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 7201 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 7261 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 7321 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 7381 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 7441 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 7501 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 7561 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 7621 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 7681 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 7741 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 7801 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 7861 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 7921 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 7981 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 8041 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 8101 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 8161 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 8221 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 8281 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 8341 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 8401 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 8461 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 8521 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 8581 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 8641 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 8701 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 8761 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 8821 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 8881 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 8941 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 9001 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 9061 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 9121 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 9181 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 9241 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 9301 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat 9361 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 9421 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat 9481 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta 9541 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata 9601 tgtactacaa acttagtagt ttttaaagaa attgtatttg ttaaatatgt actacaaact 9661 tagtagt //