LOCUS V024220 9439 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024220 VERSION V024220 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers promoter 9..235 /note="Rous sarcoma virus enhancer/promoter" /label="RSV promoter" LTR 236..416 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 463..588 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1081..1314 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1499..1543 /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 1810..1926 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2130..3312 /note="strong constitutive promoter for human elongation factor EF-1α" /label="EF-1α promoter" promoter 3329..3347 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 3369..4262 /label="AZGP1(NM_001185)" /note="AZGP1(NM_001185)" /gene="AZGP1" misc_feature 4271..4844 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES" CDS 4878..5276 /codon_start=1 /gene="Aspergillus terreus BSD" /note="confers resistance to blasticidin" /product="blasticidin S deaminase" /transl_table=1 /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG*" /label="BSD" misc_feature 5291..5879 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5966..6199 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6271..6392 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6432..6567 /note="SV40 origin of replication" /label="SV40 ori" promoter complement(6588..6606) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(6616..6632) /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" rep_origin 6774..7229 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 7255..7359 /gene="bla" /label="AmpR promoter" CDS 7360..8220 /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" rep_origin 8391..8979 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" protein_bind 9267..9288 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 9303..9333 /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 9341..9357 /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" primer_bind 9365..9381 /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" promoter 9402..9420 /note="promoter for bacteriophage T3 RNA polymerase" /label="T3 promoter" ORIGIN 1 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta 61 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg 121 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac 181 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc 241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg 421 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact 481 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa 541 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg 601 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat 661 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc 721 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag 781 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat 841 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac 901 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat 961 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg 1021 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat 1081 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat 1141 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt 1201 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca 1261 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat 1321 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag 1381 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat 1441 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa 1501 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat 1561 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt 1621 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt 1681 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga 1741 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta 1801 tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt 1861 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa 1921 aattttatcg atgcctcccc gtcaccaccc cccccaaccc gccccgaccg gagctgagag 1981 taattcatac aaaaggactc gcccctgcct tggggaatcc cagggaccgt cgttaaactc 2041 ccactaacgt agaacccaga gatcgctgcg ttcccgcccc ctcacccgcc cgctctcgtc 2101 atcactgagg tggagaagag catgcgtgag gctccggtgc ccgtcagtgg gcagagcgca 2161 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga 2221 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg 2281 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg 2341 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta 2401 cgggttatgg cccttgcgtg ccttgaatta cttccacgcc cctggctgca gtacgtgatt 2461 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga 2521 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga 2581 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat 2641 ttttgatgat atcctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa 2701 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc 2761 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg 2821 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg 2881 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt 2941 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt 3001 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc 3061 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg 3121 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag 3181 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag 3241 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt 3301 caggtgtcgt gaaaactacc cctctagata atacgactca ctatagggcc cgggttggat 3361 ccgccaccat ggtaagaatg gtgcctgtcc tgctgtctct gctgctgctt ctgggtcctg 3421 ctgtccccca ggagaaccaa gatggtcgtt actctctgac ctatatctac actgggctgt 3481 ccaagcatgt tgaagacgtc cccgcgtttc aggcccttgg ctcactcaat gacctccagt 3541 tctttagata caacagtaaa gacaggaagt ctcagcccat gggactctgg agacaggtgg 3601 aaggaatgga ggattggaag caggacagcc aacttcagaa ggccagggag gacatcttta 3661 tggagaccct gaaagacatc gtggagtatt acaacgacag taacgggtct cacgtattgc 3721 agggaaggtt tggttgtgag atcgagaata acagaagcag cggagcattc tggaaatatt 3781 actatgatgg aaaggactac attgaattca acaaagaaat cccagcctgg gtccccttcg 3841 acccagcagc ccagataacc aagcagaagt gggaggcaga accagtctac gtgcagcggg 3901 ccaaggctta cctggaggag gagtgccctg cgactctgcg gaaatacctg aaatacagca 3961 aaaatatcct ggaccggcaa gatcctccct ctgtggtggt caccagccac caggccccag 4021 gagaaaagaa gaaactgaag tgcctggcct acgacttcta cccagggaaa attgatgtgc 4081 actggactcg ggccggcgag gtgcaggagc ctgagttacg gggagatgtt cttcacaatg 4141 gaaatggcac ttaccagtcc tgggtggtgg tggcagtgcc cccgcaggac acagccccct 4201 actcctgcca cgtgcagcac agcagcctgg cccagcccct cgtggtgccc tgggaggcca 4261 gctaggctag cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat 4321 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg 4381 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc 4441 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt 4501 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg 4561 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac 4621 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg 4681 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg 4741 ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc 4801 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacactagag 4861 atccaccggt cgccaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa 4921 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag 4981 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac 5041 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt 5101 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac 5161 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc 5221 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagcgg 5281 ccgcgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 5341 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 5401 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 5461 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 5521 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 5581 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 5641 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc 5701 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 5761 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 5821 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 5881 tggaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 5941 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat 6001 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct 6061 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta 6121 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca 6181 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt 6241 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac 6301 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 6361 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc 6421 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 6481 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 6541 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc 6601 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 6661 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 6721 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc 6781 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 6841 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 6901 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 6961 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 7021 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 7081 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 7141 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 7201 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga 7261 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 7321 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 7381 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 7441 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 7501 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 7561 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 7621 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 7681 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 7741 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 7801 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 7861 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 7921 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 7981 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 8041 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 8101 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 8161 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 8221 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 8281 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 8341 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 8401 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 8461 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 8521 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 8581 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 8641 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 8701 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 8761 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 8821 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 8881 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 8941 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 9001 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 9061 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 9121 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg 9181 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg 9241 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag 9301 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt 9361 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg 9421 gaacaaaagc tggagctgc //