LOCUS V040928 9553 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040928 VERSION V040928 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..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2303..3661 /label="GPR39(NM_001508)" /note="GPR39(NM_001508)" /gene="GPR39" promoter 3712..3923 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3936..4204 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4233..4242 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4263..4925 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 4995..5048 /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 5049..5648 /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 5649..6236 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6310..6543 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6615..6736 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6755..6890 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6928..6944) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6952..6968 /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(6976..7006) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7021..7042 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7330..7918) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8089..8949) /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(8950..9054) /gene="bla" /label="AmpR promoter" primer_bind 9528..9544 /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 attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc gaattcgcca ccatggcttc acccagcctc ccgggcagtg actgctccca 2341 aatcattgat cacagtcatg tccccgagtt tgaggtggcc acctggatca aaatcaccct 2401 tattctggtg tacctgatca tcttcgtgat gggccttctg gggaacagcg ccaccattcg 2461 ggtcacccag gtgctgcaga agaaaggata cttgcagaag gaggtgacag accacatggt 2521 gagtttggct tgctcggaca tcttggtgtt cctcatcggc atgcccatgg agttctacag 2581 catcatctgg aatcccctga ccacgtccag ctacaccctg tcctgcaagc tgcacacttt 2641 cctcttcgag gcctgcagct acgctacgct gctgcacgtg ctgacactca gctttgagcg 2701 ctacatcgcc atctgtcacc ccttcaggta caaggctgtg tcgggacctt gccaggtgaa 2761 gctgctgatt ggcttcgtct gggtcacctc cgccctggtg gcactgccct tgctgtttgc 2821 catgggtact gagtaccccc tggtgaacgt gcccagccac cggggtctca cttgcaaccg 2881 ctccagcacc cgccaccacg agcagcccga gacctccaat atgtccatct gtaccaacct 2941 ctccagccgc tggaccgtgt tccagtccag catcttcggc gccttcgtgg tctacctcgt 3001 ggtcctgctc tccgtagcct tcatgtgctg gaacatgatg caggtgctca tgaaaagcca 3061 gaagggctcg ctggccgggg gcacgcggcc tccgcagctg aggaagtccg agagcgaaga 3121 gagcaggacc gccaggaggc agaccatcat cttcctgagg ctgattgttg tgacattggc 3181 cgtatgctgg atgcccaacc agattcggag gatcatggct gcggccaaac ccaagcacga 3241 ctggacgagg tcctacttcc gggcgtacat gatcctcctc cccttctcgg agacgttttt 3301 ctacctcagc tcggtcatca acccgctcct gtacacggtg tcctcgcagc agtttcggcg 3361 ggtgttcgtg caggtgctgt gctgccgcct gtcgctgcag cacgccaacc acgagaagcg 3421 cctgcgcgta catgcgcact ccaccaccga cagcgcccgc tttgtgcagc gcccgttgct 3481 cttcgcgtcc cggcgccagt cctctgcaag gagaactgag aagattttct taagcacttt 3541 tcagagcgag gccgagcccc agtctaagtc ccagtcattg agtctcgagt cactagagcc 3601 caactcaggc gcgaaaccag ccaattctgc tgcagagaat ggttttcagg agcatgaagt 3661 ttgaggatcc gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc 3721 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct 3781 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc 3841 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca 3901 acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg 3961 cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg 4021 cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac 4081 cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt 4141 gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca 4201 gatccaagct gtgaccggcg cctacgctag acgccaccat ggagagcgac gagagcggcc 4261 tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg gagttcgagc 4321 tggtgggcgg cggagagggc acccccaagc agggccgcat gaccaacaag atgaagagca 4381 ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc tacggcttct 4441 accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc atcaacaacg 4501 gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg cacgtgagct 4561 tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtggtg ggcaccggct 4621 tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc accgtggagc 4681 acctgcaccc catgggcgat aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc 4741 gcgacggcgg ctactacagc ttcgtggtgg acagccacat gcacttcaag agcgccatcc 4801 accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc 4861 acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc cccatcgcct 4921 tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct 4981 ccaccggatc tcgcgagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc 5041 ccggccctat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca 5101 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg 5161 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg 5221 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca 5281 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt 5341 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc 5401 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg 5461 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg 5521 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg 5581 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg 5641 gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 5701 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 5761 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 5821 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 5881 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 5941 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 6001 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat 6061 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 6121 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 6181 cgcgtctccg ccttcgccct cagacgagtc ggatctccct ttggccgcct ccccgcctgg 6241 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 6301 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta 6361 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 6421 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 6481 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 6541 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 6601 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 6661 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 6721 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc 6781 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 6841 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg 6901 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 6961 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 7021 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7081 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7141 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7201 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7261 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7321 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7381 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7441 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7501 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7561 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7621 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7681 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7741 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7801 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7861 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7921 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7981 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 8041 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8101 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8161 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8221 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8281 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8341 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8401 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8461 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8521 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8581 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8641 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8701 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8761 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8821 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8881 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8941 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 9001 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9061 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9121 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9181 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9241 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9301 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9361 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9421 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9481 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9541 cagtgccaag ctg //