LOCUS V043727 8894 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043727 VERSION V043727 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..3813 /label="SHMT2(NM_005412)" /note="SHMT2(NM_005412)" /gene="SHMT2" promoter 3864..4075 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4088..4356 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4383..4982 /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 4989..5577 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5651..5884 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5956..6077 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6096..6231 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6269..6285) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6293..6309 /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(6317..6347) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6362..6383 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6671..7259) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7430..8290) /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(8291..8395) /gene="bla" /label="AmpR promoter" primer_bind 8869..8885 /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 catgctgtac ttctctttgt tttgggcggc tcggcctctg 2341 cagagatgtg ggcagctggt caggatggcc attcgggctc agcacagcaa cgcagcccag 2401 actcagactg gggaagcaaa caggggctgg acaggccagg agagcctgtc ggacagtgat 2461 cctgagatgt gggagttgct gcagagggag aaggacaggc agtgtcgtgg cctggagctc 2521 attgcctcag agaacttctg cagccgagct gcgctggagg ccctggggtc ctgtctgaac 2581 aacaagtact cggagggtta tcctggcaag agatactatg ggggagcaga ggtggtggat 2641 gaaattgagc tgctgtgcca gcgccgggcc ttggaagcct ttgacctgga tcctgcacag 2701 tggggagtca atgtccagcc ctactccggg tccccagcca acctggccgt ctacacagcc 2761 cttctgcaac ctcacgaccg gatcatgggg ctggacctgc ccgatggggg ccatctcacc 2821 cacggctaca tgtctgacgt caagcggata tcagccacgt ccatcttctt cgagtctatg 2881 ccctataagc tcaaccccaa aactggcctc attgactaca accagctggc actgactgct 2941 cgacttttcc ggccacggct catcatagct ggcaccagcg cctatgctcg cctcattgac 3001 tacgcccgca tgagagaggt gtgtgatgaa gtcaaagcac acctgctggc agacatggcc 3061 cacatcagtg gcctggtggc tgccaaggtg attccctcgc ctttcaagca cgcggacatc 3121 gtcaccacca ctactcacaa gactcttcga ggggccaggt cagggctcat cttctaccgg 3181 aaaggggtga aggctgtgga ccccaagact ggccgggaga tcccttacac atttgaggac 3241 cgaatcaact ttgccgtgtt cccatccctg caggggggcc cccacaatca tgccattgct 3301 gcagtagctg tggccctaaa gcaggcctgc acccccatgt tccgggagta ctccctgcag 3361 gttctgaaga atgctcgggc catggcagat gccctgctag agcgaggcta ctcactggta 3421 tcaggtggta ctgacaacca cctggtgctg gtggacctgc ggcccaaggg cctggatgga 3481 gctcgggctg agcgggtgct agagcttgta tccatcactg ccaacaagaa cacctgtcct 3541 ggagaccgaa gtgccatcac accgggcggc ctgcggcttg gggccccagc cttaacttct 3601 cgacagttcc gtgaggatga cttccggaga gttgtggact ttatagatga aggggtcaac 3661 attggcttag aggtgaagag caagactgcc aagctccagg atttcaaatc cttcctgctt 3721 aaggactcag aaacaagtca gcgtctggcc aacctcaggc aacgggtgga gcagtttgcc 3781 agggccttcc ccatgcctgg ttttgatgag cattgaggat ccgcggccgc gaaggatctg 3841 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt 3901 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga 3961 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag 4021 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa 4081 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc 4141 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc 4201 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag 4261 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt 4321 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct 4381 agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg 4441 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg 4501 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg 4561 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg 4621 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg 4681 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg 4741 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg 4801 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc 4861 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg 4921 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct 4981 gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 5041 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 5101 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 5161 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 5221 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 5281 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 5341 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt 5401 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 5461 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 5521 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg 5581 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa 5641 aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt 5701 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 5761 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 5821 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 5881 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata 5941 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc 6001 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 6061 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt 6121 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 6181 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt 6241 gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc 6301 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 6361 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 6421 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 6481 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 6541 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 6601 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 6661 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 6721 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 6781 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 6841 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 6901 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 6961 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 7021 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 7081 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 7141 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 7201 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 7261 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 7321 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 7381 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 7441 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 7501 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 7561 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 7621 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 7681 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 7741 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 7801 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 7861 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 7921 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 7981 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 8041 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 8101 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 8161 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 8221 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 8281 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 8341 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 8401 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata 8461 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc 8521 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca 8581 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg 8641 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat 8701 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg 8761 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg 8821 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg 8881 ccagtgccaa gctg //