LOCUS V018905 9702 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018905 VERSION V018905 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2827..4398 /label="SLC2A2(NM_000340)" /note="SLC2A2(NM_000340)" /gene="SLC2A2" CDS 4405..4470 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4490..4989 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5010..5609 /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 5623..6211 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6418..7051 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7180..7196) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7204..7220 /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(7228..7258) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7273..7294 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7582..8170) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8341..9201) /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(9202..9306) /gene="bla" /label="AmpR promoter" polyA_signal 9354..9488 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 gccaccatga cagaagataa ggtcactggg accctggttt tcactgtcat cactgctgtg 2881 ctgggttcct tccagtttgg atatgacatt ggtgtgatca atgcacctca acaggtaata 2941 atatctcact atagacatgt tttgggtgtt ccactggatg accgaaaagc tatcaacaac 3001 tatgttatca acagtacaga tgaactgccc acaatctcat actcaatgaa cccaaaacca 3061 accccttggg ctgaggaaga gactgtggca gctgctcaac taatcaccat gctctggtcc 3121 ctgtctgtat ccagctttgc agttggtgga atgactgcat cattctttgg tgggtggctt 3181 ggggacacac ttggaagaat caaagccatg ttagtagcaa acattctgtc attagttgga 3241 gctctcttga tggggttttc aaaattggga ccatctcata tacttataat tgctggaaga 3301 agcatatcag gactatattg tgggctaatt tcaggcctgg ttcctatgta tatcggtgaa 3361 attgctccaa ccgctctcag gggagcactt ggcacttttc atcagctggc catcgtcacg 3421 ggcattctta ttagtcagat tattggtctt gaatttatct tgggcaatta tgatctgtgg 3481 cacatcctgc ttggcctgtc tggtgtgcga gccatccttc agtctctgct actctttttc 3541 tgtccagaaa gccccagata cctttacatc aagttagatg aggaagtcaa agcaaaacaa 3601 agcttgaaaa gactcagagg atatgatgat gtcaccaaag atattaatga aatgagaaaa 3661 gaaagagaag aagcatcgag tgagcagaaa gtctctataa ttcagctctt caccaattcc 3721 agctaccgac agcctattct agtggcactg atgctgcatg tggctcagca attttccgga 3781 atcaatggca ttttttacta ctcaaccagc atttttcaga cggctggtat cagcaaacct 3841 gtttatgcaa ccattggagt tggcgctgta aacatggttt tcactgctgt ctctgtattc 3901 cttgtggaga aggcagggcg acgttctctc tttctaattg gaatgagtgg gatgtttgtt 3961 tgtgccatct tcatgtcagt gggacttgtg ctgctgaata agttctcttg gatgagttat 4021 gtgagcatga tagccatctt cctctttgtc agcttctttg aaattgggcc aggcccgatc 4081 ccctggttca tggtggctga gtttttcagt caaggaccac gtcctgctgc tttagcaata 4141 gctgcattca gcaattggac ctgcaatttc attgtagctc tgtgtttcca gtacattgcg 4201 gacttctgtg gaccttatgt gtttttcctc tttgctggag tgctcctggc ctttaccctg 4261 ttcacatttt ttaaagttcc agaaaccaaa ggaaagtctt ttgaggaaat tgctgcagaa 4321 ttccaaaaga agagtggctc agcccacagg ccaaaagctg ctgtagaaat gaaattccta 4381 ggagctacag agactgtggg atccgactac aaagaccatg acggtgatta taaagatcat 4441 gacatcgact acaaggatga cgatgacaag tagtctagat aattctaccg ggtaggggag 4501 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc 4561 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct 4621 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc 4681 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt 4741 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc 4801 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg 4861 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg 4921 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg 4981 ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg 5041 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc 5101 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag 5161 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg 5221 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga 5281 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag 5341 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct 5401 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg 5461 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct 5521 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct 5581 ggtgcatgac ccgcaagccc ggtgcctgac cgcgtctgga acaatcaacc tctggattac 5641 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 5701 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 5761 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 5821 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 5881 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 5941 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 6001 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg 6061 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6121 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6181 agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt cgagacctag 6241 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc 6301 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac 6361 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg 6421 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac 6481 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga 6541 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata 6601 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga 6661 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc 6721 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg 6781 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct 6841 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc 6901 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga 6961 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga 7021 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag 7081 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgctagcgt 7141 tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 7201 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 7261 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 7321 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 7381 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 7441 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 7501 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 7561 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7621 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7681 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7741 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7801 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7861 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7921 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7981 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 8041 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 8101 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 8161 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 8221 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 8281 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 8341 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 8401 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 8461 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 8521 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 8581 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 8641 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 8701 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 8761 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 8821 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 8881 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8941 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 9001 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 9061 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 9121 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 9181 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 9241 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 9301 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt 9361 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 9421 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 9481 tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc 9541 ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat 9601 tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagttttta 9661 aagaaattgt atttgttaaa tatgtactac aaacttagta gt //