LOCUS V024135 9242 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024135 VERSION V024135 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..3285 /label="AKR1C3(NM_003739)" /note="AKR1C3(NM_003739)" /gene="AKR1C3" CDS 3292..3357 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3401..3612 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3625..3893 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3922..3931 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3952..4614 /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 4684..4737 /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 4738..5337 /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 5338..5925 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5999..6232 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6304..6425 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6444..6579 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6617..6633) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6641..6657 /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(6665..6695) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6710..6731 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7019..7607) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7778..8638) /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(8639..8743) /gene="bla" /label="AmpR promoter" primer_bind 9217..9233 /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 attccaaaca ccagtgtgta 2341 aagctaaatg atggccactt catgcctgta ttgggatttg gcacctatgc acctccagag 2401 gttccgagaa gtaaagcttt ggaggtcaca aaattagcaa tagaagctgg gttccgccat 2461 atagattctg ctcatttata caataatgag gagcaggttg gactggccat ccgaagcaag 2521 attgcagatg gcagtgtgaa gagagaagac atattctaca cttcaaagct ttggtccact 2581 tttcatcgac cagagttggt ccgaccagcc ttggaaaact cactgaagaa agctcaattg 2641 gactatgttg acctctatct tattcattct ccaatgtctc taaagccagg tgaggaactt 2701 tcaccaacag atgaaaatgg aaaagtaata tttgacatag tggatctctg taccacctgg 2761 gaggccatgg agaagtgtaa ggatgcagga ttggccaagt ccattggggt gtcaaacttc 2821 aaccgcaggc agctggagat gatcctcaac aagccaggac tcaagtacaa gcctgtctgc 2881 aaccaggtag aatgtcatcc gtatttcaac cggagtaaat tgctagattt ctgcaagtcg 2941 aaagatattg ttctggttgc ctatagtgct ctgggatctc aacgagacaa acgatgggtg 3001 gacccgaact ccccggtgct cttggaggac ccagtccttt gtgccttggc aaaaaagcac 3061 aagcgaaccc cagccctgat tgccctgcgc taccagctgc agcgtggggt tgtggtcctg 3121 gccaagagct acaatgagca gcgcatcaga cagaacgtgc aggtttttga gttccagttg 3181 actgcagagg acatgaaagc catagatggc ctagacagaa atctccacta ttttaacagt 3241 gatagttttg ctagccaccc taattatcca tattcagatg aatatggatc cgactacaaa 3301 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 3361 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 3421 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 3481 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 3541 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 3601 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 3661 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 3721 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 3781 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 3841 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 3901 tgaccggcgc ctacgctaga cgccaccatg gagagcgacg agagcggcct gcccgccatg 3961 gagatcgagt gccgcatcac cggcaccctg aacggcgtgg agttcgagct ggtgggcggc 4021 ggagagggca cccccaagca gggccgcatg accaacaaga tgaagagcac caaaggcgcc 4081 ctgaccttca gcccctacct gctgagccac gtgatgggct acggcttcta ccacttcggc 4141 acctacccca gcggctacga gaaccccttc ctgcacgcca tcaacaacgg cggctacacc 4201 aacacccgca tcgagaagta cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc 4261 tacgaggccg gccgcgtgat cggcgacttc aaggtggtgg gcaccggctt ccccgaggac 4321 agcgtgatct tcaccgacaa gatcatccgc agcaacgcca ccgtggagca cctgcacccc 4381 atgggcgata acgtgctggt gggcagcttc gcccgcacct tcagcctgcg cgacggcggc 4441 tactacagct tcgtggtgga cagccacatg cacttcaaga gcgccatcca ccccagcatc 4501 ctgcagaacg ggggccccat gttcgccttc cgccgcgtgg aggagctgca cagcaacacc 4561 gagctgggca tcgtggagta ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc 4621 cgcgctcagt cgtccaattc tgccgtggac ggcaccgccg gacccggctc caccggatct 4681 cgcgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg 4741 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 4801 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 4861 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 4921 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 4981 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 5041 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 5101 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 5161 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 5221 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 5281 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat 5341 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 5401 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 5461 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 5521 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 5581 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 5641 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 5701 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc 5761 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 5821 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtctccgc 5881 cttcgccctc agacgagtcg gatctccctt tggccgcctc cccgcctggt acctttaaga 5941 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 6001 gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc 6061 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 6121 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 6181 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt 6241 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag 6301 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 6361 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 6421 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc 6481 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 6541 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc 6601 caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 6661 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 6721 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 6781 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 6841 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 6901 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 6961 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 7021 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 7081 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 7141 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 7201 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 7261 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 7321 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 7381 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 7441 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 7501 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 7561 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 7621 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 7681 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 7741 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 7801 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 7861 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 7921 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 7981 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 8041 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 8101 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 8161 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 8221 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 8281 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 8341 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 8401 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 8461 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 8521 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 8581 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 8641 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 8701 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 8761 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 8821 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 8881 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 8941 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 9001 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 9061 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 9121 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 9181 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc 9241 tg //