LOCUS V042700 9064 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042700 VERSION V042700 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..3918 /label="ALDH7A1(NM_001182)" /note="ALDH7A1(NM_001182)" /gene="ALDH7A1" CDS 3925..3990 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4034..4245 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4258..4526 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4553..5152 /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 5159..5747 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5821..6054 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6126..6247 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6266..6401 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6439..6455) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6463..6479 /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(6487..6517) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6532..6553 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6841..7429) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7600..8460) /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(8461..8565) /gene="bla" /label="AmpR promoter" primer_bind 9039..9055 /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 catgtggcgc cttcctcgcg cgctgtgtgt gcacgctgca 2341 aagaccagca agctctctgg accttggagc aggcctgccg ccttcatgtc cactctcctc 2401 atcaatcagc cccagtatgc gtggctgaaa gagctggggc tccgcgagga aaacgagggc 2461 gtgtataatg gaagctgggg aggccgggga gaggttatta cgacctattg ccctgctaac 2521 aacgagccaa tagcaagagt ccgacaggcc agtgtggcag actatgaaga aactgtaaag 2581 aaagcaagag aagcatggaa aatctgggca gatattcctg ctccaaaacg aggagaaata 2641 gtaagacaga ttggcgatgc cttgcgggag aagatccaag tactaggaag cttggtgtct 2701 ttggagatgg ggaaaatctt agtggaaggt gtgggtgaag ttcaggagta tgtggatatc 2761 tgtgactatg ctgttggttt atcaaggatg attggaggac ctatcttgcc ttctgaaaga 2821 tctggccatg cactgattga gcagtggaat cccgtaggcc tggttggaat catcacggca 2881 ttcaatttcc ctgtggcagt gtatggttgg aacaacgcca tcgccatgat ctgtggaaat 2941 gtctgcctct ggaaaggagc tccaaccact tccctcatta gtgtggctgt cacaaagata 3001 atagccaagg ttctggagga caacaagctg cctggtgcaa tttgttcctt gacttgtggt 3061 ggagcagata ttggcacagc aatggccaaa gatgaacgag tgaacctgct gtccttcact 3121 gggagcactc aggtgggaaa acaggtgggc ctgatggtgc aggagaggtt tgggagaagt 3181 ctgttggaac ttggaggaaa caatgccatt attgcctttg aagatgcaga cctcagctta 3241 gttgttccat cagctctctt cgctgctgtg ggaacagctg gccagaggtg taccactgcg 3301 aggcgactgt ttatacatga aagcatccat gatgaggttg taaacagact taaaaaggcc 3361 tatgcacaga tccgagttgg gaacccatgg gaccctaatg ttctctatgg gccactccac 3421 accaagcagg cagtgagcat gtttcttgga gcagtggaag aagcaaagaa agaaggtggc 3481 acagtggtct atgggggcaa ggttatggat cgccctggaa attatgtaga accgacaatt 3541 gtgacaggtc ttggccacga tgcgtccatt gcacacacag agacttttgc tccgattctc 3601 tatgtcttta aattcaagaa tgaagaagag gtctttgcat ggaataatga agtaaaacag 3661 ggactttcaa gtagcatctt taccaaagat ctgggcagaa tctttcgctg gcttggacct 3721 aaaggatcag actgtggcat tgtaaatgtc aacattccaa caagtggggc tgagattgga 3781 ggtgcctttg gaggagaaaa gcacactggt ggtggcaggg agtctggcag tgatgcctgg 3841 aaacagtaca tgagaaggtc tacttgtact atcaactaca gtaaagacct tcctctggcc 3901 caaggaatca agtttcaggg atccgactac aaagaccatg acggtgatta taaagatcat 3961 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc 4021 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 4081 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 4141 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 4201 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 4261 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 4321 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 4381 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 4441 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 4501 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 4561 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 4621 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 4681 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4741 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4801 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4861 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4921 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 4981 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 5041 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 5101 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 5161 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 5221 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 5281 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5341 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5401 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5461 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5521 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 5581 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5641 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5701 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 5761 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5821 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5881 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5941 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 6001 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 6061 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6121 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6181 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6241 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6301 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6361 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6421 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6481 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6541 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6601 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6661 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6721 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6781 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6841 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6901 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6961 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7021 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7081 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7141 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7201 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7261 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7321 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7381 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7441 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7501 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7561 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7621 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7681 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7741 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7801 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7861 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7921 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7981 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8041 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8101 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8161 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8221 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8281 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8341 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8401 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8461 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8521 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8581 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8641 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8701 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8761 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8821 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8881 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8941 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9001 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9061 gctg //