LOCUS V033206 9562 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033206 VERSION V033206 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 2843..4318 /label="SLC2A1(NM_006516)" /note="SLC2A1(NM_006516)" /gene="SLC2A1" promoter 4350..4849 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4870..5469 /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 5483..6071 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6278..6911 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7040..7056) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7064..7080 /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(7088..7118) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7133..7154 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7442..8030) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8201..9061) /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(9062..9166) /gene="bla" /label="AmpR promoter" polyA_signal 9214..9348 /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 ctcaagcttc gaattcgcca ccatggagcc cagcagcaag aagctgacgg gtcgcctcat 2881 gctggccgtg ggaggagcag tgcttggctc cctgcagttt ggctacaaca ctggagtcat 2941 caatgccccc cagaaggtga tcgaggagtt ctacaaccag acatgggtcc accgctatgg 3001 ggagagcatc ctgcccacca cgctcaccac gctctggtcc ctctcagtgg ccatcttttc 3061 tgttgggggc atgattggct ccttctctgt gggccttttc gttaaccgct ttggccggcg 3121 gaattcaatg ctgatgatga acctgctggc cttcgtgtcc gccgtgctca tgggcttctc 3181 gaaactgggc aagtcctttg agatgctgat cctgggccgc ttcatcatcg gtgtgtactg 3241 cggcctgacc acaggcttcg tgcccatgta tgtgggtgaa gtgtcaccca cagcccttcg 3301 tggggccctg ggcaccctgc accagctggg catcgtcgtc ggcatcctca tcgcccaggt 3361 gttcggcctg gactccatca tgggcaacaa ggacctgtgg cccctgctgc tgagcatcat 3421 cttcatcccg gccctgctgc agtgcatcgt gctgcccttc tgccccgaga gtccccgctt 3481 cctgctcatc aaccgcaacg aggagaaccg ggccaagagt gtgctaaaga agctgcgcgg 3541 gacagctgac gtgacccatg acctgcagga gatgaaggaa gagagtcggc agatgatgcg 3601 ggagaagaag gtcaccatcc tggagctgtt ccgctccccc gcctaccgcc agcccatcct 3661 catcgctgtg gtgctgcagc tgtcccagca gctgtctggc atcaacgctg tcttctatta 3721 ctccacgagc atcttcgaga aggcgggggt gcagcagcct gtgtatgcca ccattggctc 3781 cggtatcgtc aacacggcct tcactgtcgt gtcgctgttt gtggtggagc gagcaggccg 3841 gcggaccctg cacctcatag gcctcgctgg catggcgggt tgtgccatac tcatgaccat 3901 cgcgctagca ctgctggagc agctaccctg gatgtcctat ctgagcatcg tggccatctt 3961 tggctttgtg gccttctttg aagtgggtcc tggccccatc ccatggttca tcgtggctga 4021 actcttcagc cagggtccac gtccagctgc cattgccgtt gcaggcttct ccaactggac 4081 ctcaaatttc attgtgggca tgtgcttcca gtatgtggag caactgtgtg gtccctacgt 4141 cttcatcatc ttcactgtgc tcctggttct gttcttcatc ttcacctact tcaaagttcc 4201 tgagactaaa ggccggacct tcgatgagat cgcttccggc ttccggcagg ggggagccag 4261 ccaaagtgac aagacacccg aggagctgtt ccatcccctg ggggctgatt cccaagtgtg 4321 aggatcccgc gactctagat aattctaccg ggtaggggag gcgcttttcc caaggcagtc 4381 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc 4441 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt 4501 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc 4561 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac 4621 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt 4681 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct 4741 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa 4801 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc 4861 aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 4921 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 4981 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 5041 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 5101 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 5161 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 5221 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 5281 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 5341 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 5401 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 5461 ggtgcctgac cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac 5521 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 5581 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 5641 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 5701 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 5761 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 5821 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct 5881 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 5941 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 6001 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 6061 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca 6121 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag 6181 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct 6241 gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa 6301 cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattag 6361 cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag 6421 ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg 6481 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg 6541 tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag 6601 aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt 6661 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt 6721 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 6781 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 6841 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 6901 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa 6961 atgaatatca gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg acctctagct 7021 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7081 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7141 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7201 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7261 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7321 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7381 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7441 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7501 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7561 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7621 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7681 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7741 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7801 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7861 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 7921 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7981 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8041 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8101 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8161 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8221 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8281 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8341 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8401 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8461 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8521 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8581 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8641 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8701 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8761 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8821 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8881 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8941 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9001 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9061 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9121 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9181 aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc ttataatggt 9241 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 9301 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact 9361 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac 9421 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat 9481 tgtatttgtt aaatatgtac tacaaactta gtagttttta aagaaattgt atttgttaaa 9541 tatgtactac aaacttagta gt //