LOCUS V032951 9478 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032951 VERSION V032951 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..3631 /label="Bscl2(NM_001136064)" /note="Bscl2(NM_001136064)" /gene="Bscl2" promoter 3676..3887 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3900..4168 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4203..4910 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" /label="mCherry" CDS 4920..4973 /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 4974..5573 /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 5574..6161 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6235..6468 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6540..6661 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6680..6815 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6853..6869) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6877..6893 /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(6901..6931) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6946..6967 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7255..7843) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8014..8874) /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(8875..8979) /gene="bla" /label="AmpR promoter" primer_bind 9453..9469 /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 ccatgataca tcaaagaaga gaagctgggg caagagagac 2341 atgcagagac cagatcaaag gatcagacaa agacgaggaa ccttcagctg ccctatccca 2401 tggccaaggg tatcgtccct gtggaagacc agccaggaac tcaaagcctg aggctggggc 2461 cagaccccct gctgtcccca tcatggtcaa tgacccacca gtccccgcct tactgtgggc 2521 ccaggaagtg ggccacgtct tggcaggccg tgcccgcagg ctgatgctgc agtttggggt 2581 gctcttctgc accatccttc ttttgctttg ggtgtctgtg ttcctctatg gctccttcta 2641 ctactcctac atgccgacgg tcagccacct cagccctgtg cacttccact acaggacaga 2701 ctgtgattcc tccaccgcct ctctctgctc tttccctgtt gccaatgtct cactggctaa 2761 gagtggacgt gatcgggtac tgatgtatgg acagccatat cgagtcacct tagagcttga 2821 actgcccgaa tcccctgtga atcaagactt gggcatgttc ttagtcactg tttcatgtta 2881 taccagaggt ggccgaatca tctccacttc ttcacgctca gtgatgctgc attaccgctc 2941 gcagctgctc caggtgctcg acacgctgct gttctccagc ctcctgctgt ttggcttcgc 3001 tgaacagaag cagctcctgg aagtagaact ctactctgac tatagggaga attcgtatgt 3061 gccaacaact ggagcaatta ttgagatcca cagcaagcgc atccagatgt atggagccta 3121 cctccggatc catgcccact tcactgggct caggtacctg ctgtacaact tccccatgac 3181 ctgcgccttc gtgggtgtgg ccagcaactt cacgttcctc agcgtcatcg tgctcttcag 3241 ctatatgcag tgggtgtggg gtgctgtctg gccccgccac cgcttctctc tgcaggttaa 3301 tatccgacaa agggataact cacaccacgg ggccccgcgt cggatctcgc gccatcagcc 3361 aggtcaggaa tctacccagc agtcggatgt gacggaggat ggtgagagcc ctgaagatcc 3421 ctcaggaaca gagggccagc tgtctgagga agagaaacca gagaagcggc ccctgaatgg 3481 agaggaggag caagagccag aggctagtga tggctcctgg gaagatgcag ctttgctgac 3541 agaggccaac ccacctacct ctgcctcagc atctgccctt gcccctgaga ctctgggtag 3601 cctcaggcaa cgcccgacct gctccagttc ctgagcggcc gcgaaggatc tgcgatcgct 3661 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 3721 gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 3781 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 3841 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag 3901 gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt 3961 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg 4021 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta 4081 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt 4141 cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctagacgcca 4201 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca 4261 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg 4321 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc 4381 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc 4441 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc 4501 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg 4561 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg 4621 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg 4681 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg 4741 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg 4801 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg 4861 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag ggatctcgcg 4921 agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc cctatgaccg 4981 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc 5041 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca 5101 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca 5161 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg 5221 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc 5281 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt 5341 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg 5401 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct 5461 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg 5521 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac 5581 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 5641 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 5701 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 5761 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 5821 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 5881 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 5941 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg 6001 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 6061 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt ctccgccttc 6121 gccctcagac gagtcggatc tccctttggc cgcctccccg cctggtacct ttaagaccaa 6181 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag 6241 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt 6301 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 6361 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 6421 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat 6481 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga 6541 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 6601 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 6661 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc 6721 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct 6781 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa 6841 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6901 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6961 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 7021 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 7081 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 7141 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 7201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 7261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7681 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7801 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7861 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7921 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7981 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 8041 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 8101 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 8161 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 8221 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 8281 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8341 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8401 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8461 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8521 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8581 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8641 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8701 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8761 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8821 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8881 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8941 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 9001 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 9061 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 9121 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 9181 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 9241 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9301 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9361 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9421 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //