LOCUS V033941 8905 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033941 VERSION V033941 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /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 1804..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3759 /label="CELF4(NM_020180)" /note="CELF4(NM_020180)" /gene="CELF4" CDS 3766..3831 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3875..4086 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4099..4367 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4394..4993 /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 5000..5588 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5662..5895 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5967..6088 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6107..6242 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6280..6296) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6304..6320 /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(6328..6358) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6373..6394 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6682..7270) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7441..8301) /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(8302..8406) /gene="bla" /label="AmpR promoter" primer_bind 8880..8896 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgtatata aagatggcca cgttagcaaa cggacaggct 2341 gacaacgcaa gcctcagtac caacgggctc ggcagcagcc cgggcagtgc cgggcacatg 2401 aacggattaa gccacagccc ggggaacccg tcgaccattc ccatgaagga ccacgatgcc 2461 atcaagctgt tcattgggca gatcccccgc aacctggatg agaaggacct caagcccctc 2521 ttcgaggagt ttggcaaaat ctacgagctt acggttctga aggacaggtt cacaggcatg 2581 cacaaaggct gcgccttcct cacctactgc gagcgtgagt cagcgctgaa ggcccagagc 2641 gcgctgcacg agcagaagac tctgcccggg atgaaccggc cgatccaggt gaagcctgcg 2701 gacagcgaga gccgaggagg tagtagctgc ctgcgccagc ccccttcaca agatagaaaa 2761 ctcttcgtgg gcatgctcaa caagcaacag tccgaggacg acgtgcgccg ccttttcgag 2821 gcctttggga acatcgagga gtgcaccatc ctgcgcgggc ccgacggcaa cagcaagggg 2881 tgcgcctttg tgaagtactc ctcccacgcc gaggcgcagg ccgccatcaa cgcgctacac 2941 ggcagccaga ccatgccggg agcctcgtcc agtctggtgg tcaagttcgc cgacaccgac 3001 aaggagcgca cgatgcggcg aatgcagcag atggctggcc agatgggcat gttcaacccc 3061 atggccatcc ctttcggggc ctacggcgcc tacgctcagg cactgatgca gcagcaagcg 3121 gccctgatgg catcagtcgc gcagggcggc tacctgaacc ccatggctgc cttcgctgcc 3181 gcccagatgc agcagatggc ggccctcaac atgaatggcc tggcggccgc acctatgacc 3241 ccaacctcag gtggcagcac ccctccgggc atcactgcac cagccgtgcc tagcatccca 3301 tcccccattg gggtgaatgg cttcaccggc ctccccccac aggccaatgg gcaacctgct 3361 gcggaagctg tgttcgccaa tggcatccac ccctacccag cacagagccc caccgccgcg 3421 gaccccctgc agcaggccta cgccggagtg cagcagtatg caggtccagc tgcctaccct 3481 gctgcctatg gtcagataag ccaggccttt cctcagccgc ctccaatgat cccccagcag 3541 cagagagaag ggcccgaggg ctgtaacctg ttcatctacc atctgcccca ggagtttggg 3601 gacgctgagc tgatgcagat gttcctccct ttcggcttcg tgagcttcga caacccggcc 3661 agcgcgcaga ccgccatcca ggccatgaac ggcttccaga tcggcatgaa gaggctcaag 3721 gtgcagctga agcggcccaa agacgccaat cgcccgtacg gatccgacta caaagaccat 3781 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3841 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 3901 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3961 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 4021 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 4081 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 4141 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 4201 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 4261 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 4321 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4381 gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt 4441 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac 4501 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg 4561 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg 4621 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc 4681 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca 4741 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg 4801 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt 4861 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt 4921 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa 4981 gcccggtgcc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg 5041 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 5101 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 5161 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 5221 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 5281 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 5341 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc 5401 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 5461 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 5521 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc 5581 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 5641 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc 5701 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 5761 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 5821 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 5881 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa 5941 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat 6001 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 6061 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac 6121 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 6181 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 6241 cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg 6301 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 6361 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 6421 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 6481 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 6541 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 6601 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 6661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 6721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 6781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 6841 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 6901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 6961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7081 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 7141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7201 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 7261 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 7321 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 7381 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 7441 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 7501 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 7561 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 7621 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 7681 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 7741 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 7801 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 7861 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 7921 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 7981 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8041 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 8101 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 8161 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 8221 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 8281 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 8341 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 8401 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 8461 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 8521 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 8581 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 8641 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 8701 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 8761 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 8821 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 8881 taaaacgacg gccagtgcca agctg //