LOCUS V029018 8223 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029018 VERSION V029018 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 65..368 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 369..572 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" protein_bind 595..619 /bound_moiety="BP Clonase™" /gene="mutant version of attB" /note="recombination site for the Gateway® BP reaction" /label="attB1" CDS 619..1122 /label="DESI1(NM_015704)" /note="DESI1(NM_015704)" /gene="DESI1" protein_bind complement(1121..1145) /bound_moiety="BP Clonase™" /gene="mutant version of attB" /note="recombination site for the Gateway® BP reaction" /label="attB2" CDS 1147..1188 /codon_start=1 /product="epitope tag from simian virus 5" /transl_table=1 /translation="GKPIPNPLLGLDST" /label="V5 tag" misc_feature 1230..1818 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 1890..2123 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 2201..2322 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 2362..2497 /note="SV40 origin of replication" /label="SV40 ori" promoter complement(2518..2536) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(2546..2562) /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" rep_origin 2704..3159 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3185..3289 /gene="bla" /label="AmpR promoter" CDS 3290..4150 /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 4321..4909 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" protein_bind 5197..5218 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 5233..5263 /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5271..5287 /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 5295..5311 /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" promoter 5332..5350 /note="promoter for bacteriophage T3 RNA polymerase" /label="T3 promoter" promoter 5378..5604 /note="Rous sarcoma virus enhancer/promoter" /label="RSV promoter" LTR 5605..5785 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 5832..5957 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 6450..6683 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 6868..6912 /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" promoter 7087..7587 /note="human phosphoglycerate kinase 1 promoter" /label="hPGK promoter" CDS 7599..7994 /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 8054..8171 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" ORIGIN 1 cccggggtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt 61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc 121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac 181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata 241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc 301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta 361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac 421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc 481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc 541 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactgtcggg atcaacaagt 601 ttgtacaaaa aagttggcat ggagccgccg aatctctatc cggtgaagct ctacgtgtac 661 gacctgtcca aaggcctggc ccggcggctc agccccatca tgctggggaa acaactggaa 721 ggcatctggc acacatccat agttgtgcac aaggatgagt tcttcttcgg cagtggtggt 781 atctccagct gccccccggg agggacattg cttgggcctc cagactctgt ggttgatgtg 841 gggagtacag aagtcacaga agaaatcttt ctggagtacc tctcctccct gggggagtcc 901 ctgttccgag gtgaggccta caacctcttt gaacacaatt gtaacacctt cagcaacgaa 961 gtggcacagt tcctgactgg gcggaagatt ccttcttaca tcacagacct gccctctgaa 1021 gttctctcca cgccctttgg acaggcactt cggcccctcc tggactccat tcagatccag 1081 cctccaggag ggagctccgt gggcagaccc aacggccaga gcccaacttt cttgtacaaa 1141 gtggttggta agcctatccc taaccctctc ctcggtctcg attctacgta gtaatgagct 1201 agcgctaacc ggtggcgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa 1261 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 1321 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 1381 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 1441 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 1501 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 1561 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 1621 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga 1681 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 1741 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 1801 tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct 1861 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca 1921 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct 1981 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca 2041 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta 2101 gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta 2161 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta 2221 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 2281 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct 2341 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 2401 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 2461 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct 2521 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa 2581 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta 2641 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat 2701 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 2761 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 2821 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 2881 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 2941 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 3001 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 3061 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 3121 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa 3181 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 3241 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca 3301 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 3361 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 3421 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 3481 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 3541 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 3601 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 3661 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 3721 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 3781 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 3841 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 3901 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 3961 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat 4021 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 4081 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 4141 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 4201 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 4261 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 4321 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 4381 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 4441 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 4501 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 4561 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 4621 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 4681 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 4741 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 4801 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 4861 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 4921 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 4981 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 5041 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 5101 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 5161 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 5221 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg 5281 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc 5341 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg 5401 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac 5461 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag 5521 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt 5581 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag 5641 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 5701 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 5761 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga 5821 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg 5881 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag 5941 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc 6001 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg 6061 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa 6121 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata 6181 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag 6241 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg 6301 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa 6361 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg 6421 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca 6481 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg 6541 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg 6601 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga 6661 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc 6721 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac 6781 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta 6841 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa 6901 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc 6961 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg 7021 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg 7081 ggacccttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg 7141 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt 7201 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct 7261 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa 7321 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc 7381 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc 7441 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc 7501 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt 7561 gaccgaatca ccgacctctc tccccagggg gtaccaccat ggccaagcct ttgtctcaag 7621 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag 7681 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg 7741 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg 7801 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga 7861 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag 7921 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt 7981 atgtgtggga gggcctgcag ctgcagtagt aagaattcta gatcttgaga caaatggcag 8041 tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa 8101 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa 8161 ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggcgc cggctcgagg 8221 ggg //