LOCUS pLV3-CMV-ZDHHC6( 9531 bp DNA circular SYN 15-OCT-2026 DEFINITION synthetic circular DNA. ACCESSION V044508 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9531) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..9531 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 233..413 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 460..585 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1078..1311 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1496..1540 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 1807..1923 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2013..2216 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 2319..3557 /gene="ZDHHC6" /label=ZDHHC6(NM_022494) /note="ZDHHC6(NM_022494)" CDS 3564..3629 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" promoter 3673..3884 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 3897..4165 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" regulatory 4194..4203 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 4224..4886 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /label=CopGFP /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" CDS 4956..5009 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 5010..5609 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" misc_feature 5610..6198 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 6272..6505 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 6577..6698 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 6738..6873 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(6906..6922) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 6930..6946 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6954..6984) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6999..7020 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(7308..7896) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8067..8927) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8928..9032) /gene="bla" /label=AmpR promoter primer_bind 9506..9522 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" 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 cgatacataa acgggtctct 241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc 421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg 481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt 541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg 601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa 661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg 721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca 781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa 841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa 901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag 961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt 1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg 1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac 1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct 1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct 1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg 1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa 1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa 1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa 1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac 1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag 1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc 1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga 1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg 1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga 1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat 1921 tttatcgata ctagtattat gcccagtaca tgaccttatg ggactttcct acttggcagt 1981 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg 2041 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg 2101 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc 2161 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ttatataagc agagctcgtt 2221 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagat 2281 tctagagcta gcctgatacg aactcggaat tcgccaccat gggtacattc tgttcggtta 2341 tcaagtttga aaatctacaa gaattaaaga gactgtgtca ctggggtccc atcatagccc 2401 ttggtgttat agcaatatgt tctaccatgg ccatgattga ctctgtgttg tggtattggc 2461 ccttacatac aactggagga agtgtgaatt tcatcatgtt gataaattgg actgtcatga 2521 ttctttataa ttacttcaat gccatgtttg tcggtccggg ctttgtccct ctggggtgga 2581 aaccggaaat ttctcaggat accatgtatc tccagtattg taaagtctgc caagcataca 2641 aggcaccacg ttcacatcac tgcagaaagt gtaacagatg tgtgatgaag atggaccatc 2701 actgtccttg gatcaacaac tgttgtggtt accaaaatca tgcttcgttc acactgtttc 2761 tccttttagc accactgggt tgtatccatg ctgctttcat ttttgtgatg actatgtaca 2821 cacagcttta tcatcggctc tcctttgggt ggaacacagt gaagatcgac atgagtgcag 2881 cccggagaga tcctcttcca attgttccat ttggattagc tgcatttgct accaccttgt 2941 ttgccttggg attagcttta ggaacaacca tagctgttgg gatgttgttt tttatccaga 3001 tgaaaataat tctcagaaac aaaacttcta ttgagtcatg gattgaagag aaggctaaag 3061 atcgaattca gtattatcaa ctagatgaag tctttgtttt tccatatgat atgggaagta 3121 gatggaggaa ctttaaacag gtatttacgt ggtcaggggt ccctgaagga gatggacttg 3181 agtggccagt aagagaaggc tgtcaccaat acagcttaac aatagaacag ttgaaacaaa 3241 aagcagataa gagagtcaga agtgttcgct ataaagtaat agaagattat agtggtgcct 3301 gctgccctct gaataaagga atcaaaacct tcttcacaag tccctgcacc gaagagcctc 3361 gaatacagct gcaaaaaggg gaattcattt tagccacaag aggtttacga tactggttat 3421 atggagacaa aattcttgat gattccttta tagaaggtgt ttcaagaata aggggttggt 3481 tccctagaaa atgtgtggaa aagtgtccct gtgatgctga aacagatcaa gccccagagg 3541 gggagaagaa aaatagagga tccgactaca aagaccatga cggtgattat aaagatcatg 3601 acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc gatcgctccg 3661 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg 3721 tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg 3781 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg 3841 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg 3901 ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga 3961 gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa 4021 gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac 4081 tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt 4141 tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta gacgccacca 4201 tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc accggcaccc 4261 tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag cagggccgca 4321 tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac ctgctgagcc 4381 acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac gagaacccct 4441 tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag tacgaggacg 4501 gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg atcggcgact 4561 tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac aagatcatcc 4621 gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg gtgggcagct 4681 tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg gacagccaca 4741 tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc atgttcgcct 4801 tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag taccagcacg 4861 ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg 4921 acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt cttctaacat 4981 gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg 5041 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc 5101 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag 5161 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg 5221 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga 5281 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag 5341 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct 5401 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg 5461 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct 5521 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct 5581 ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa 5641 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 5701 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 5761 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 5821 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 5881 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 5941 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 6001 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga 6061 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 6121 tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctccc 6181 tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat 6241 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga 6301 taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag 6361 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 6421 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 6481 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta 6541 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat 6601 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 6661 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc 6721 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 6781 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat 6841 tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac caaattcgta 6901 atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 6961 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 7021 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 7081 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 7141 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 7201 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 7261 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 7321 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 7381 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 7441 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 7501 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 7561 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 7621 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 7681 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 7741 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 7801 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 7861 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 7921 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 7981 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 8041 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 8101 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 8161 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 8221 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 8281 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 8341 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 8401 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 8461 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 8521 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 8581 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 8641 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 8701 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 8761 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 8821 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 8881 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 8941 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 9001 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 9061 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc 9121 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 9181 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 9241 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg 9301 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 9361 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 9421 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg 9481 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct g //