LOCUS V034939 10030 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034939 VERSION V034939 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 2317..4056 /label="Slc5a7(NM_022025)" /note="Slc5a7(NM_022025)" /gene="Slc5a7" CDS 4063..4128 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4172..4383 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4396..4664 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4693..4702 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4723..5385 /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 5455..5508 /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 5509..6108 /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 6109..6697 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6771..7004 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7076..7197 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7237..7372 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7405..7421) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7429..7445 /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(7453..7483) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7498..7519 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7807..8395) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8566..9426) /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(9427..9531) /gene="bla" /label="AmpR promoter" primer_bind 10005..10021 /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 ctgatacgaa ctcggaattc gccaccatgt ctttccacgt agaaggactg 2341 gtagctatta tcctcttcta cctccttata tttctggttg gaatatgggc tgcatggaaa 2401 accaaaaaca gcggcaaccc agaagagcgc agtgaagcca tcatagtcgg gggccgtgac 2461 attggtttgt tggttggtgg ttttaccatg acagccacct gggttggagg aggctacatc 2521 aatgggacag cagaagcagt gtatgggcca ggttgtggtc tagcttgggc tcaggcaccc 2581 attggatatt ctctgagtct aattttaggt ggtctgtttt ttgcgaaacc tatgcgttcc 2641 aagggatatg tgactatgtt agacccattt caacagatct atggaaagcg catgggtggg 2701 ctgctcttca tccctgcact gatgggagag atgttctggg ctgcagcaat tttctctgca 2761 ttaggggcca ccatcagcgt gatcattgat gtggatgtga acatatcggt cattgtctct 2821 gcactcattg ccattcttta taccctagtg ggtgggctct actctgtggc atatactgat 2881 gttgtccagc tattctgcat ttttatagga ctgtggatca gtgtcccttt tgccctgtca 2941 catcctgcag tcaccgacat cggattcaca gctgtgcatg ctaaatacca gagtccctgg 3001 ctgggaacca ttgaatcagt tgaagtctac acctggcttg ataattttct gttattgatg 3061 ctgggtggaa tcccatggca agcctacttc cagagggtcc tctcttcatc ctcagccacc 3121 tatgctcagg tactgtcctt cctggcagct tttgggtgcc tggtgatggc tctacccgcc 3181 atatgcatag gagctattgg agcttccaca gactggaacc agactgccta cgggtatcca 3241 gatcccaaga ctaaggagga agcagacatg attctcccga tcgttctgca gtacctctgc 3301 cctgtgtaca tctccttctt tgggcttggt gctgtttcag ctgctgtcat gtcctcagct 3361 gactcgtcca tcctgtcggc gagttctatg tttgctcgga atatctacca gctttccttc 3421 agacaaaatg catcagacaa ggaaattgtg tgggtcatga ggatcactgt gcttgtgttc 3481 ggagcatctg caacagccat ggctttgctg acgaagactg tgtatgggct ctggtacctg 3541 agctctgacc ttgtctacat catcatcttc ccacagctgc tctgtgtact cttcatcaaa 3601 ggaaccaaca cttatggggc agttgctggt tatatttttg gactattcct gagaattact 3661 ggaggagagc catatctata cttgcagccc ttaatcttct accctggtta ttactctgac 3721 aagaatggta tatacaatca gaggttccca tttaaaactc tctccatggt tacctcattc 3781 tttaccaaca tttgtgtttc ttatctagcc aagtatctat ttgaaagtgg aaccttgcct 3841 ccaaaattag atgtatttga tgctgttgtc gcaaggcaca gtgaagagaa catggacaag 3901 accattctag tcagaaatga aaatatcaaa ttaaatgaac ttgcacctgt gaaacctcgg 3961 cagagcctaa ccctcagttc aactttcacc aataaggagg ccctccttga tgttgattcc 4021 agtccggagg ggtctgggac tgaagataac ttacaaggat ccgactacaa agaccatgac 4081 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 4141 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 4201 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 4261 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 4321 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 4381 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 4441 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 4501 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 4561 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 4621 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 4681 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 4741 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 4801 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 4861 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 4921 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 4981 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 5041 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 5101 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 5161 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 5221 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 5281 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 5341 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 5401 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 5461 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 5521 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 5581 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 5641 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 5701 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5761 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5821 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5881 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5941 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 6001 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 6061 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 6121 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 6181 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 6241 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 6301 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 6361 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 6421 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 6481 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 6541 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 6601 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 6661 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 6721 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 6781 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 6841 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6901 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6961 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 7021 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 7081 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 7141 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 7201 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 7261 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 7321 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 7381 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 7441 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 7501 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 7561 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 7621 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7681 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7741 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7801 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7861 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7921 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7981 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 8041 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 8101 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 8161 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 8221 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 8281 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 8341 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 8401 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 8461 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 8521 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 8581 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 8641 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 8701 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8761 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8821 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8881 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8941 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 9001 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 9061 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 9121 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 9181 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 9241 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 9301 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 9361 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 9421 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 9481 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 9541 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 9601 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 9661 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 9721 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9781 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9841 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9901 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9961 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 10021 tgccaagctg //