LOCUS V037871 9002 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037871 VERSION V037871 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" regulatory 2288..2297 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2297..2362 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2369..3904 /label="Slc16a14(NM_027921)" /note="Slc16a14(NM_027921)" /gene="Slc16a14" promoter 3972..4183 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4196..4464 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4491..5090 /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 5097..5685 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5759..5992 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6064..6185 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6204..6339 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6377..6393) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6401..6417 /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(6425..6455) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6470..6491 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6779..7367) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7538..8398) /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(8399..8503) /gene="bla" /label="AmpR promoter" primer_bind 8977..8993 /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga 2341 ctacaaggat gacgatgaca aggaattcat gtatacgagt catgaagata tcgggtatga 2401 ccttgaagac gaccgaaagg ctaagaataa gaagacgctg aagccccacc cagatatcga 2461 tggcgggtgg gcctggatga tggtcctctc gtccttcttt gttcacatcc tcatcatggg 2521 ctcacagatg gccctaggag tcctcaacgt ggagtggctt gaagagtttc accagagccg 2581 cggcctgacc gcctgggtca gctccctgag catgggcatc accttgatcg tgggaccttt 2641 catcggcttg ttcattaaca cctgcgggtg ccgccagaca gcaatcattg gaggcctggt 2701 gaactccctc ggctgggtgt tgagcgccta tgcagccaat gtgcagtctc tctttattac 2761 ctttggagtg gcagcaggcc ttggcagtgg gatggcttac ctacctgcag tggtcatggt 2821 gggaaggtac tttcagaaga gacgagccct ggcccagggc ctcagtacca cggggacagg 2881 atttgggacg ttcctgatga ccgtgttgct gaaatacctg tgcgcagaat atggctggcg 2941 gaatgctatg ttcatccaag gtgctctgtc cctgaacctg tgtgtctgtg gggcgctcat 3001 gaggcccctc tctcccgaga agttagaaaa ctgcccagaa gcggaagagc cgtgtgctct 3061 cccagcttac tccactgaat ctgttaagtc tggaggacca ctgggcatgg ctgaagaaca 3121 ggacagaagg cccgggaatg aggagatggt gtgtgacctt caaacacagg agtgtcaagg 3181 tcaaacccat ccgaggaaga atgtgtgtgc cttccgggtt ctgaagacag tgagccagct 3241 cactgtgcaa gtccggaggg gctttaggga ctggcactca ggctattttg ggacggcttc 3301 actcttcacc aaccgcatgt ttgtagcctt tattttctgg gctctgttcg catacagcag 3361 ctttgtcatc cctttcatcc acctgcccga aatcgtcagt ttgtataact tgtcagagca 3421 aaatgacaca ttccctctga cctcgattat agcaatactt cacatcttcg ggaaggtgat 3481 cttgggggcc gtggccgacc tcccctgtat cagcgtctgg aatgtcttcc tcatcgctaa 3541 cttcaccctc gtcctcagca tcttccttct gcctctgatg cacacctacg cgagtctggc 3601 tgtcatttgt gccctaattg ggttttctag cgggtatttc tccctcatgc ctgtggtgac 3661 agaggacctg gttggcactg agcacttggc caatgcctac ggcatcatca tctgcgccaa 3721 tggcatctca gccttactag gaccaccatt cgcaggatgg atctttgaca tcacccaaaa 3781 atatgacttt tccttttata tatgtggttt gctttacatg gtagggatac tctttttgct 3841 cattcaaccg tgtattcaga tgatagatca atccagaagg aagtgcatag agggtgcaca 3901 tgtttaggga tccctcgaga gatcagtgta gcggccgcga aggatctgcg atcgctccgg 3961 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt 4021 cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt 4081 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc 4141 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc 4201 tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag 4261 tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag 4321 tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact 4381 cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt 4441 ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt 4501 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg 4561 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg 4621 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg 4681 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag 4741 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg 4801 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt 4861 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg 4921 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg 4981 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg 5041 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc 5101 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 5161 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 5221 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 5281 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 5341 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 5401 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 5461 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct 5521 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 5581 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 5641 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga 5701 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 5761 gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc 5821 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 5881 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 5941 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt 6001 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag 6061 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 6121 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 6181 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc 6241 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 6301 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc 6361 caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 6421 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 6481 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 6541 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 6601 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 6661 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 6721 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 6781 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 6841 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 6901 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 6961 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 7021 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 7081 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 7141 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 7201 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 7261 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 7321 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 7381 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 7441 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 7501 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 7561 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 7621 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 7681 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 7741 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 7801 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 7861 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 7921 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 7981 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 8041 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 8101 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 8161 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 8221 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 8281 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 8341 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 8401 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 8461 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 8521 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 8581 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 8641 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 8701 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 8761 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 8821 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 8881 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 8941 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc 9001 tg //