LOCUS V036044 9310 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036044 VERSION V036044 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..3336 /label="ST3GAL1(NM_003033)" /note="ST3GAL1(NM_003033)" /gene="ST3GAL1" CDS 3343..3408 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3452..3663 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3676..3944 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3973..3982 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4003..4665 /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 4735..4788 /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 4789..5388 /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 5389..5977 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6051..6284 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6356..6477 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6517..6652 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6685..6701) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6709..6725 /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(6733..6763) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6778..6799 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7087..7675) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7846..8706) /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(8707..8811) /gene="bla" /label="AmpR promoter" primer_bind 9285..9301 /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 gccaccatgg tgaccctgcg gaagaggacc 2341 ctgaaagtgc tcaccttcct cgtgctcttc atcttcctca cctccttctt cctgaactac 2401 tcccacacca tggtggccac cacctggttc cccaagcaga tggtcctgga gctctccgag 2461 aacctgaaga gactgatcaa gcacaggcct tgcacctgca cccactgcat cgggcagcgc 2521 aagctctcgg cctggttcga tgagaggttc aaccagacca tgcagccgct gctgaccgcc 2581 cagaacgcgc tcttggagga cgacacctac cgatggtggc tgaggctcca gcgggagaag 2641 aagcccaata acttgaatga caccatcaag gagctgttca gagtggtgcc tgggaatgtg 2701 gaccctatgc tggagaagag gtcggtgggc tgccggcgct gcgccgttgt gggcaactcg 2761 ggcaacctga gggagtcttc ttatgggcct gagatagaca gtcacgactt tgtcctcagg 2821 atgaacaagg cgcccacggc agggtttgaa gctgatgttg ggaccaagac cacccaccat 2881 ctggtgtacc ctgagagctt ccgggagctg ggagataatg tcagcatgat cctggtgccc 2941 ttcaagacca tcgacttgga gtgggtggtg agcgccatca ccacgggcac catttcccac 3001 acctacatcc cggttcctgc aaagatcaga gtgaaacagg ataagatcct gatctaccac 3061 ccagccttca tcaagtatgt ctttgacaac tggctgcaag ggcacgggcg atacccatct 3121 accggcatcc tctcggtcat cttctcaatg catgtctgcg atgaggtgga cttgtacggc 3181 ttcggggcag acagcaaagg gaactggcac cactactggg agaacaaccc atccgcgggg 3241 gcttttcgca agacgggggt gcacgatgca gactttgagt ctaacgtgac ggccaccttg 3301 gcctccatca ataaaatccg gatcttcaag gggagaggat ccgactacaa agaccatgac 3361 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 3421 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 3481 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 3541 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 3601 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 3661 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 3721 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 3781 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 3841 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 3901 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 3961 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 4021 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 4081 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 4141 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 4201 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 4261 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 4321 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4381 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 4441 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 4501 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 4561 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 4621 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 4681 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 4741 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 4801 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 4861 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 4921 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 4981 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5041 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5101 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5161 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5221 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5281 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5341 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 5401 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5461 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5521 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 5581 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5641 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 5701 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 5761 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 5821 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 5881 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 5941 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 6001 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 6061 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 6121 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6181 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6241 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6301 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6361 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6421 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6481 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 6541 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 6601 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 6661 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 6721 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6781 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6841 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6901 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6961 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7021 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7081 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7141 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7201 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7261 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7321 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7381 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7441 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7501 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7561 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7621 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7681 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7741 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7801 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7861 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7921 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7981 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8041 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8101 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8161 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8221 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8281 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8341 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8401 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8461 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8521 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8581 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8641 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8701 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8761 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8821 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8881 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8941 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 9001 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9061 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9121 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9181 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9241 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9301 tgccaagctg //