LOCUS V044563 8503 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044563 VERSION V044563 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..3357 /label="Wnt3a(NM_009522)" /note="Wnt3a(NM_009522)" /gene="Wnt3a" CDS 3364..3429 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3473..3684 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3697..3965 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3992..4591 /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 4598..5186 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5260..5493 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5565..5686 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5705..5840 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5878..5894) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5902..5918 /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(5926..5956) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5971..5992 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6280..6868) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7039..7899) /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(7900..8004) /gene="bla" /label="AmpR promoter" primer_bind 8478..8494 /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 catggctcct ctcggatacc tcttagtgct ctgcagcctg 2341 aagcaggctc tgggcagcta cccgatctgg tggtccttgg ctgtgggacc ccagtactcc 2401 tctctgagca ctcagcccat tctctgtgcc agcatcccag gcctggtacc gaagcagctg 2461 cgcttctgca ggaactacgt ggagatcatg cccagcgtgg ctgagggtgt caaagcgggc 2521 atccaggagt gccagcacca gttccgaggc cggcgttgga actgcaccac cgtcagcaac 2581 agcctggcca tctttggccc tgttctggac aaagccaccc gggagtcagc ctttgtccat 2641 gccatcgcct ccgctggagt agctttcgca gtgacacgct cctgtgcaga gggatcagct 2701 gctatctgtg ggtgcagcag ccgcctccag ggctccccag gcgagggctg gaagtggggc 2761 ggctgtagtg aggacattga atttggagga atggtctctc gggagtttgc cgatgccagg 2821 gagaaccggc cggatgcccg ctctgccatg aaccgtcaca acaatgaggc tgggcgccag 2881 gccatcgcca gtcacatgca cctcaagtgc aaatgccacg ggctatctgg cagctgtgaa 2941 gtgaagacct gctggtggtc gcagccggac ttccgcacca tcggggattt cctcaaggac 3001 aagtatgaca gtgcctcgga gatggtggta gagaaacacc gagagtctcg tggctgggtg 3061 gagaccctga ggccacgtta cacgtacttc aaggtgccga cagaacgcga cctggtctac 3121 tacgaggcct cacccaactt ctgcgaacct aaccccgaaa ccggctcctt cgggacgcgt 3181 gaccgcacct gcaatgtgag ctcgcatggc atagatgggt gcgacctgtt gtgctgcggg 3241 cgcgggcata acgcgcgcac tgagcgacgg agggagaaat gccactgtgt tttccattgg 3301 tgctgctacg tcagctgcca ggagtgcaca cgtgtctatg acgtgcacac ctgcaaggga 3361 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 3421 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 3481 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 3541 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 3601 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 3661 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 3721 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 3781 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 3841 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 3901 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 3961 agatccaagc tgtgaccggc gcctacgcta gatgaccgag tacaagccca cggtgcgcct 4021 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta 4081 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca 4141 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg 4201 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga 4261 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga 4321 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt 4381 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc 4441 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt 4501 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac 4561 ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat caacctctgg attacaaaat 4621 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 4681 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 4741 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 4801 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 4861 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 4921 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 4981 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct 5041 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 5101 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 5161 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag 5221 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc 5281 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag 5341 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 5401 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 5461 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc 5521 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag 5581 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 5641 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct 5701 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 5761 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg 5821 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc 5881 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 5941 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 6001 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 6061 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 6121 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 6181 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 6241 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 6301 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 6361 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 6421 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 6481 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 6541 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 6601 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 6661 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 6721 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 6781 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 6841 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 6901 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 6961 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 7021 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 7081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 7141 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 7201 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 7261 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 7321 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 7381 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 7441 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 7501 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 7561 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 7621 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 7681 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 7741 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 7801 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 7861 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 7921 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 7981 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga 8041 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct 8101 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac 8161 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt 8221 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca 8281 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca 8341 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt 8401 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt 8461 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg //