LOCUS V027107 9744 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027107 VERSION V027107 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 1494..1538 /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 1801..1916 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 1974..2277 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2278..2481 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2588..3814 /label="Neu1(NM_010893)" /note="Neu1(NM_010893)" /gene="Neu1" CDS 3839..3910 /codon_start=1 /product="three tandem FLAG® epitope tags" /transl_table=1 /translation="DYKDDDDKDYKDDDDKDYKDDDDK" /label="3xFLAG" promoter 3964..4175 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4188..4456 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4484..5176 /codon_start=1 /note="mammalian codon-optimized" /product="Zoanthus green fluorescent protein" /transl_table=1 /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH AIASGSALP" /label="ZsGreen1" CDS 5186..5239 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 5240..5839 /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 5840..6427 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6501..6734 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6806..6927 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6946..7081 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7119..7135) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7143..7159 /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(7167..7197) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7212..7233 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7521..8109) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8280..9140) /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(9141..9245) /gene="bla" /label="AmpR promoter" primer_bind 9719..9735 /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 gggcgcagcg 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 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca 1441 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg 1501 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa 1561 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa 1621 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt 1681 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag 1741 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgcc 1801 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata 1861 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg 1921 gtttattaca gggacagcag agatccagtt tatcgataag cttgggagtt ccgcgttaca 1981 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 2041 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 2101 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 2161 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 2221 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 2281 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 2341 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2401 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2461 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca 2521 cgctgttttg acctccatag aagacaccga ctctactaga ggatctattt ccggtgaatt 2581 cgccaccatg gtgggggcag acccgaccag accccgggga ccgctgagct attgggcggg 2641 ccgtcggggt caggggctcg cagcgatctt cctgctcctg gtgtccgcgg cggaatccga 2701 ggccagggca gaggatgact tcagcctggt gcagccgctg gtgaccatgg agcagctgct 2761 gtgggtgagc gggaagcaga tcggctctgt agacactttc cgcatcccgc tcatcacagc 2821 cacccctcgg ggcacgctcc tggccttcgc tgaggccagg aaaaaatctg catccgatga 2881 gggggccaag ttcatcgcca tgaggaggtc cacggaccag ggtagcacgt ggtcctctac 2941 agccttcatc gtagacgatg gggaggcctc cgatggcctg aacctgggcg ctgtggtgaa 3001 cgatgtagac acagggatag tgttccttat ctataccctc tgtgctcaca aggtcaactg 3061 ccaggtggcc tctaccatgt tggtttggag taaggacgac ggcatttcct ggagcccacc 3121 ccggaatctc tctgtggata ttggcacaga gatgtttgcc cctggacctg gctcaggcat 3181 tcagaaacag cgggagcctg ggaagggccg gctcattgtg tgtggacacg ggacgctgga 3241 gcgagatggg gtcttctgtc tcctcagtga tgaccacggt gcctcctggc actacggcac 3301 tggagtgagc ggcattccct ttggccagcc caaacacgat cacgatttca accccgacga 3361 gtgccagccc tacgagcttc cagatggctc ggtcatcatc aacgcccgga accagaataa 3421 ctaccattgc cgctgcagga tcgtcctccg cagctatgac gcctgtgaca ccctcaggcc 3481 ccgggatgtg accttcgacc ctgagctcgt ggaccctgtg gtagctgcag gagcactagc 3541 caccagctcc ggcattgtct tcttctccaa tccagcccac cctgagttcc gagtgaacct 3601 gaccctgcgc tggagtttca gcaatggtac atcctggcag aaggagaggg tccaggtgtg 3661 gccgggaccc agcggctact cgtccctgac agccctggaa aacagcacgg atggaaagaa 3721 gcagcccccg cagctgttcg ttctgtacga gaaaggcctg aaccggtaca ccgagagcat 3781 ctccatggtc aaaatcagcg tctacggcac gctcggatcc tcggtaccaa gcttaagtga 3841 ctacaaggat gacgatgaca aggattacaa agacgacgat gataaggact ataaggatga 3901 tgacgacaaa taaagatcca tcgatactag taaggatctg cgatcgctcc ggtgcccgtc 3961 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt 4021 gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc 4081 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg 4141 ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct 4201 ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc 4261 tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc 4321 tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc 4381 tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc 4441 tgcgccgtta cagatccaag ctgtgaccgg cgcctactct agcatggccc agtccaagca 4501 cggcctgacc aaggagatga ccatgaagta ccgcatggag ggctgcgtgg acggccacaa 4561 gttcgtgatc accggcgagg gcatcggcta ccccttcaag ggcaagcagg ccatcaacct 4621 gtgcgtggtg gagggcggcc ccttgccctt cgccgaggac atcttgtccg ccgccttcat 4681 gtacggcaac cgcgtgttca ccgagtaccc ccaggacatc gtcgactact tcaagaactc 4741 ctgccccgcc ggctacacct gggaccgctc cttcctgttc gaggacggcg ccgtgtgcat 4801 ctgcaacgcc gacatcaccg tgagcgtgga ggagaactgc atgtaccacg agtccaagtt 4861 ctacggcgtg aacttccccg ccgacggccc cgtgatgaag aagatgaccg acaactggga 4921 gccctcctgc gagaagatca tccccgtgcc caagcagggc atcttgaagg gcgacgtgag 4981 catgtacctg ctgctgaagg acggtggccg cttgcgctgc cagttcgaca ccgtgtacaa 5041 ggccaagtcc gtgccccgca agatgcccga ctggcacttc atccagcaca agctgacccg 5101 cgaggaccgc agcgacgcca agaaccagaa gtggcacctg accgagcacg ccatcgcctc 5161 cggctccgcc ttgcccgcgg ccgctgaggg cagaggaagt cttctaacat gcggtgacgt 5221 ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 5281 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 5341 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 5401 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 5461 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 5521 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 5581 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 5641 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 5701 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 5761 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 5821 ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg 5881 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 5941 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 6001 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 6061 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 6121 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 6181 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac 6241 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 6301 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 6361 gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctccc tttggccgcc 6421 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 6481 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct 6541 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 6601 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 6661 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 6721 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 6781 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata 6841 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 6901 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact 6961 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 7021 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 7081 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga 7141 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 7201 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 7261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 7321 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 7381 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 7441 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 7501 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 7561 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 7621 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 7681 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 7741 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 7801 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 7861 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 7921 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 7981 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 8041 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 8101 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 8161 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 8221 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 8281 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 8341 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 8401 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 8461 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 8521 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 8581 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 8641 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 8701 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 8761 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 8821 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 8881 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 8941 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 9001 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 9061 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 9121 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 9181 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 9241 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 9301 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 9361 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 9421 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg 9481 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 9541 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 9601 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 9661 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 9721 aaaacgacgg ccagtgccaa gctg //