LOCUS V045179 9373 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045179 VERSION V045179 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..4227 /label="Alas1(NM_001291835)" /note="Alas1(NM_001291835)" /gene="Alas1" CDS 4234..4299 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4343..4554 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4567..4835 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4862..5461 /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 5468..6056 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6130..6363 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6435..6556 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6575..6710 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6748..6764) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6772..6788 /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(6796..6826) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6841..6862 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7150..7738) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7909..8769) /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(8770..8874) /gene="bla" /label="AmpR promoter" primer_bind 9348..9364 /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 catggagact gtcgttcgcc gatgcccatt cttatcccga 2341 gtccctcagg cctttctgca gaaggcagga aagtctctgt tgttctatgc tcaaaactgc 2401 cccaagatga tggaagttgg ggccaagcca gctcctcgaa ccctgtccac atcagctgtc 2461 cactgccagc aggtcaaaga aacccctcca gccaatgaga aagagaaaac tgccaaggcc 2521 gcagtccagc aggctcctga cgagtcccag atggcacaga ctccagacgg cacacagctt 2581 ccgtctggac acccgtcccc tgctacaagc cagggctctg ggagcaagtg ccctttcctg 2641 gcagcacagc tgagccagac aggcagcagc gtcttccgca aggccagtct ggagctccag 2701 gaggacgtgc aggaaatgca tgctgtgagg aaagaggctg ctcaaagccc agtgcctccc 2761 agcctggtta atgtgaaaac cgatggagag gatccaagcc gactgctgaa gaacttccag 2821 gacatcatgc gaaagcagag gccagaaaga gtgtctcatc ttcttcagga taacttgcca 2881 aagtctgttt ccacttttca atatgatcat ttctttgaga agaaaattga tgagaaaaaa 2941 aatgatcata cctaccgagt ttttaaaacg gtgaaccgga gagcacaaat cttcccgatg 3001 gcggatgact acacggattc cctcataacc aaaaagcagg tgtcggtctg gtgcagtaat 3061 gactacctag gcatgagtcg acacccacgg gtgtgtgggg ctgtcatgga gactgtgaaa 3121 cagcatggtg ctggagctgg gggaactagg aatatttctg gaacgagcaa gttccacgta 3181 gaactggagc aggcactggc cgacctccac ggcaaagatg cggcgctctt gttctcttcc 3241 tgttttgtgg ccaacgactc caccctcttc accctggcta agatgatgcc aggctgtgaa 3301 atttactctg attccgggaa ccatgcctcc atgatccaag ggattcgcaa cagtcgagtg 3361 ccaaaatata tcttccgcca caatgacgtt aaccatctca gagaactgct gcagagatct 3421 gacccctcag tccccaagat agtagcattt gaaactgtcc attcaatgga tggagcagtg 3481 tgcccactgg aagagctgtg tgacgtggcc catgagtttg gggcaatcac gtttgtggat 3541 gaggtccatg cagtggggct ctatggagct cgaggtggag ggatcggtga tcgggatggc 3601 gtcatgccaa aaatggacat catctctggg acgcttggta aagcgttcgg ctgtgttgga 3661 ggatacattg ccagcacgag tttgctgatc gacaccgtcc gctcctacgc tgcgggcttc 3721 atcttcacca cctccttgcc accaatgctg ctggctggag ccctggagtc tgtgcggatc 3781 ctgaagagca gcgaggggcg cgcccttcgc cgccagcacc agcgaaatgt caagcttctg 3841 aggcagatgc taatggacgc tggcctcccg gtcatccact gtccgagtca catcatccct 3901 gtgcgggttg ccgatgctgc taagaacaca gagatctgtg atgagttgat gaccaggcat 3961 aatatctacg tccaggccat caattaccca acagtgcctc gcggggagga gctcctgcgg 4021 atcgccccca ccccgcatca caccccgcag atgatgaact tcttcgtgga gaagctgctg 4081 gtcacgtgga agcgagttgg gctggaactt aagccacatt cgtcagctga atgcaacttc 4141 tgcaggaggc ccctgcactt tgaagtgatg agcgaaagag agaaagccta tttctcaggc 4201 atgagcaaga tggtgtctgc ccaggccgga tccgactaca aagaccatga cggtgattat 4261 aaagatcatg acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc 4321 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 4381 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 4441 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 4501 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 4561 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 4621 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 4681 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 4741 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 4801 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 4861 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt 4921 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga 4981 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga 5041 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga 5101 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg 5161 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga 5221 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg 5281 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct 5341 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc 5401 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg 5461 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5521 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5581 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5641 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 5701 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 5761 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 5821 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg 5881 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 5941 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 6001 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 6061 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 6121 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta 6181 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 6241 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 6301 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 6361 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 6421 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 6481 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 6541 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc 6601 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 6661 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg 6721 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 6781 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6841 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6901 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6961 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7021 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7081 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7141 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7201 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7261 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7321 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7381 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7441 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7501 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7561 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7621 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7681 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7741 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7801 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7861 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7921 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7981 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8041 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8101 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8161 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8221 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8281 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8341 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8401 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8461 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8521 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8581 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8641 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8701 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8761 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8821 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8881 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8941 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9001 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9061 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9121 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9181 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9241 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9301 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9361 cagtgccaag ctg //