LOCUS V043358 9505 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043358 VERSION V043358 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..4359 /label="Tgm2(NM_009373)" /note="Tgm2(NM_009373)" /gene="Tgm2" CDS 4366..4431 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4475..4686 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4699..4967 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4994..5593 /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 5600..6188 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6262..6495 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6567..6688 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6707..6842 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6880..6896) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6904..6920 /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(6928..6958) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6973..6994 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7282..7870) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8041..8901) /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(8902..9006) /gene="bla" /label="AmpR promoter" primer_bind 9480..9496 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggcagag gagctgctcc tggagaggtg tgatttggag 2341 attcaggcca atggccgtga ccaccacacg gccgacctat gccaagagaa actggtgctg 2401 cgtcgtggtc agcgcttccg gctgactctg tacttcgagg gccgtggcta cgaggccagc 2461 gtggacagcc tcacgttcgg tgctgtgacc ggcccagatc ccagtgaaga ggcagggacc 2521 aaggcccgct tctcactgtc tgacaatgtg gaggagggat cttggtcagc ctcagtgctg 2581 gaccaacagg acaatgtcct ctcgctacag ctctgcaccc cagccaatgc tcctattggc 2641 ctgtaccgtc tcagcctaga ggcttctact ggctaccagg gctccagctt tgtgctgggc 2701 cacttcatcc tgctctacaa tgcctggtgc ccagccgatg atgtgtacct agactcagag 2761 gaggagcgac gggaatatgt ccttacgcaa cagggcttca tctaccaagg ctctgtcaag 2821 ttcatcaaga gtgtgccttg gaactttggg cagttcgagg atggaatcct ggatacctgc 2881 ctgatgctct tggatatgaa ccccaagttc ctgaagaacc gtagtcggga ctgctcacgc 2941 cgcagcagtc ccatctatgt gggccgcgtg gtgagcgcca tggtcaactg caatgatgac 3001 cagggtgtgc ttctgggccg ctgggacaac aactatgggg atggtatcag tcccatggcc 3061 tggattggca gtgtggacat tctgcggcgc tggaaggaac acggctgtca gcaagtgaag 3121 tacgggcagt gctgggtgtt tgcagcggtg gcctgcacag tgctgcggtg cctcggcatc 3181 cctaccagag tggtgaccaa ctacaactcc gcccacgacc agaacagcaa cctgctcatc 3241 gagtacttcc gaaatgagtt cggggagctg gagagcaaca agagcgagat gatctggaac 3301 ttccactgct gggtggagtc ctggatgacc aggccagacc tacagccggg ctatgagggg 3361 tggcaggcca ttgaccccac accacaggag aagagcgaag ggacatactg ttgtggccca 3421 gtctcagtgc gagccatcaa ggagggagac ctgagtacca agtatgatgc acccttcgtg 3481 tttgccgagg tcaacgctga tgtggtggac tggatccggc aggaagatgg gtctgtgctc 3541 aaatccatca accgttcctt ggtcgtgggg cagaagatca gcactaagag tgtgggccgt 3601 gatgaccggg aggacatcac ccatacctac aagtacccag aggggtcacc cgaggagagg 3661 gaagtcttca ccaaggccaa ccacctgaac aaactggcag agaaagagga gacaggggtg 3721 gccatgcgca tccgagtggg ggacagtatg agcatgggca acgacttcga cgtgtttgcc 3781 cacatcggca acgacacctc ggagactcga gagtgtcgtc tcctgctctg tgcccgcact 3841 gtcagctaca acggggtgct ggggcccgag tgtggcactg aggacatcaa cctgaccctg 3901 gatccctact ctgagaacag catcccactt cgaatcctct acgagaagta cagcgggtgc 3961 ctgacagagt caaacctcat caaggtgcgg ggccttctca tcgaaccagc tgccaacagc 4021 tacctgctgg ctgagagaga tctctacctg gagaatcccg aaatcaagat ccgggtcctg 4081 ggagaaccca agcaaaaccg caaactggtg gctgaggtgt ccctgaagaa cccactttcc 4141 gatcccctgt atgactgcat cttcactgtg gagggggctg gcctgaccaa ggagcagaag 4201 tctgtggaag tctcagaccc ggtgccagcg ggcgatttgg tcaaggcacg ggtcgacctg 4261 ttcccgactg atattggcct ccacaagctg gtggtgaact tccagtgtga caagctgaag 4321 tcggtgaagg gttaccggaa tgttatcatc ggcccggccg gatccgacta caaagaccat 4381 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 4441 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 4501 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 4561 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 4621 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 4681 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 4741 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 4801 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 4861 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 4921 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4981 gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt 5041 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac 5101 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg 5161 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg 5221 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc 5281 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca 5341 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg 5401 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt 5461 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt 5521 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa 5581 gcccggtgcc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg 5641 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 5701 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 5761 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 5821 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 5881 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 5941 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc 6001 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 6061 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 6121 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc 6181 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 6241 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc 6301 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 6361 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 6421 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 6481 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa 6541 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat 6601 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 6661 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac 6721 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 6781 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 6841 cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg 6901 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 6961 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 7021 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 7081 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 7141 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 7201 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 7261 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7321 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7381 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7441 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7501 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7561 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7621 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7681 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 7741 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7801 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 7861 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 7921 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 7981 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 8041 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 8101 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 8161 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 8221 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 8281 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 8341 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 8401 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 8461 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 8521 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 8581 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8641 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 8701 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 8761 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 8821 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 8881 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 8941 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 9001 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 9061 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 9121 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 9181 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 9241 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 9301 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 9361 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 9421 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 9481 taaaacgacg gccagtgcca agctg //