LOCUS V028621 10821 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028621 VERSION V028621 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" promoter 2337..3519 /note="strong constitutive promoter for human elongation factor EF-1α" /label="EF-1α promoter" CDS 3548..5539 /label="SLC5A1(NM_000343)" /note="SLC5A1(NM_000343)" /gene="SLC5A1" misc_feature 5571..6157 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 6177..6776 /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 6790..7378 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7585..8218 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8346..8362) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8370..8386 /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(8394..8424) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8439..8460 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8748..9336) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9507..10367) /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(10368..10472) /gene="bla" /label="AmpR promoter" polyA_signal 10520..10654 /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 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg 2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc 2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct 2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtacgaatt 3541 cgccaccatg gacagtagca cctggagccc caagaccacc gcggtcaccc ggcctgttga 3601 gacccacgag ctcattcgca atgcagccga tatctccatc atcgttatct acttcgtggt 3661 agtgatggcc gtcggactgt gggctatgtt ttccaccaat cgtgggactg ttggaggctt 3721 cttcctggca ggccgaagta tggtgtggtg gccgattgga gcctccctct ttgctagtaa 3781 cattggaagt ggccactttg tggggctggc cgggactggg gcagcttcag gcatcgccat 3841 tggaggcttt gaatggaatg ccctggtttt ggtggttgtg ctgggctggc tgtttgtccc 3901 catctatatt aaggctgggg tggtgacaat gccagagtac ctgaggaagc ggtttggagg 3961 ccagcggatc caggtctacc tttcccttct gtccctgctg ctctacattt tcaccaagat 4021 ctcggcagac atcttctcgg gggccatatt catcaatctg gccttaggcc tgaatctgta 4081 tttagccatc tttctcttat tggcaatcac tgccctttac acaattacag ggggcctggc 4141 ggcggtgatt tacacggaca ccttgcagac ggtgatcatg ctggtggggt ctttaatcct 4201 gactgggttt gcttttcacg aagtgggagg ctatgacgcc ttcatggaaa agtacatgaa 4261 agccattcca accatagtgt ctgatggcaa caccaccttt caggaaaaat gctacactcc 4321 aagggccgac tccttccaca tcttccgaga tcccctcacg ggagacctcc catggcctgg 4381 gttcatcttt gggatgtcca tccttacctt gtggtactgg tgcacagatc aggtcattgt 4441 gcagcgctgc ctctcagcca agaatatgtc tcacgtgaag ggtggctgca tcctgtgtgg 4501 gtatctaaag ctgatgccca tgttcatcat ggtgatgcca ggaatgatca gccgcattct 4561 gtacacagaa aaaattgcct gtgtcgtccc ttcagaatgt gagaaatatt gcggtaccaa 4621 ggttggctgt accaacatcg cctatccaac cttagtggtg gagctcatgc ccaatggact 4681 gcgaggcctg atgctatcag tcatgctggc ctccctcatg agctccctga cctccatctt 4741 caacagcgcc agcaccctct tcaccatgga catctacgcc aaggtccgca agagagcatc 4801 tgagaaagag ctcatgattg ccggaaggtt gtttatcctg gtgctgattg gcatcagcat 4861 cgcctgggtg cccattgtgc agtcagcaca aagtgggcaa ctcttcgatt acatccagtc 4921 catcaccagt tacttgggac cacccattgc ggctgtcttc ctgcttgcta ttttctggaa 4981 gagagtcaat gagccaggag ccttttgggg actgatccta ggacttctga ttgggatttc 5041 acgtatgatt actgagtttg cttatggaac cgggagctgc atggagccca gcaactgtcc 5101 cacgattatc tgtggggtgc actacttgta ctttgccatt atcctcttcg ccatttcttt 5161 catcaccatc gtggtcatct ccctcctcac caaacccatt ccggatgtgc atctctaccg 5221 tctgtgttgg agcctgcgca acagcaaaga ggagcgtatt gacctggatg cggaagagga 5281 gaacatccaa gaaggcccta aggagaccat tgaaatagaa acacaagttc ctgagaagaa 5341 aaaaggaatc ttcaggagag cctatgacct attttgtggg ctagagcagc acggtgcacc 5401 caagatgact gaggaagagg agaaagccat gaagatgaag atgacggaca cctctgagaa 5461 gcctttgtgg aggacagtgt tgaacgtcaa tggcatcatc ctggtgaccg tggctgtctt 5521 ttgccatgca tattttgcct gaactagttc tagagcggcc gcggatcccg cccctctccc 5581 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc 5641 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc 5701 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt 5761 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct 5821 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa 5881 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt 5941 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag 6001 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt 6061 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt 6121 ttcctttgaa aaacacgatg ataagcttgc cacaacccac aaggagacga ccttccatga 6181 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca 6241 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc 6301 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 6361 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 6421 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 6481 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 6541 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 6601 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 6661 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 6721 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctagacgc 6781 gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 6841 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 6901 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 6961 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 7021 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 7081 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 7141 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 7201 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 7261 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 7321 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 7381 ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag 7441 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc 7501 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc 7561 actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata 7621 tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag aactacacac 7681 cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg 7741 agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga 7801 gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc 7861 tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc 7921 gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga 7981 ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg 8041 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 8101 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 8161 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 8221 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 8281 agtgagaggc cttgacattg ctagcgttta ccgtcgacct ctagctagag cttggcgtaa 8341 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 8401 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 8461 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 8521 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 8581 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 8641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 8701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 8761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 8821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 8881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 8941 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 9001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 9061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 9121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 9181 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 9241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 9301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 9361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 9421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 9481 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 9541 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 9601 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 9661 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 9721 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 9781 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 9841 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 9901 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 9961 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 10021 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 10081 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 10141 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 10201 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 10261 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 10321 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 10381 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 10441 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 10501 gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa 10561 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 10621 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa 10681 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt 10741 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat 10801 atgtactaca aacttagtag t //