LOCUS V040965 9124 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040965 VERSION V040965 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..3978 /label="GALNT1(NM_020474)" /note="GALNT1(NM_020474)" /gene="GALNT1" CDS 3985..4050 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4094..4305 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4318..4586 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4613..5212 /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 5219..5807 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5881..6114 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6186..6307 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6326..6461 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6499..6515) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6523..6539 /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(6547..6577) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6592..6613 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6901..7489) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7660..8520) /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(8521..8625) /gene="bla" /label="AmpR promoter" primer_bind 9099..9115 /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 catgagaaaa tttgcatact gcaaggtggt cctagccacc 2341 tccttgattt gggtactctt ggatatgttc ctgctgcttt acttcagtga atgcaacaaa 2401 tgtgatgaaa aaaaggagag aggacttcct gctggagatg ttctagagcc agtacaaaag 2461 cctcatgaag gtcctggaga aatggggaaa ccagtcgtca ttcctaaaga ggatcaagaa 2521 aagatgaaag agatgtttaa aatcaatcag ttcaatttaa tggcaagtga gatgattgca 2581 ctcaacagat ctttaccaga tgttaggtta gaagggtgta aaacaaaggt gtatccagat 2641 aatcttccta caacaagtgt ggtgattgtt ttccacaatg aggcttggag cacacttctg 2701 cgaactgtcc atagtgtcat taatcgctca ccaagacaca tgatagaaga aattgttcta 2761 gtagatgatg ccagtgaaag agactttttg aaaaggcctt tagagagtta tgtgaaaaaa 2821 ctaaaagtac cagttcatgt aattcgaatg gaacaacgtt ctggattgat cagagctaga 2881 ttaaaaggag ctgctgtgtc taaaggccaa gtgatcacct tcctggatgc ccattgtgag 2941 tgtacagtgg gatggctgga gcctctcttg gccaggatca aacatgacag gagaacagtg 3001 gtgtgtccca tcatcgatgt gatcagtgat gatacttttg agtacatggc aggctctgat 3061 atgacctatg gtgggttcaa ctggaagctc aattttcgct ggtatcctgt tccccaaaga 3121 gaaatggaca gaaggaaagg tgatcggact cttcctgtca ggacacctac catggcagga 3181 ggcctttttt caatagacag agattacttt caggaaattg gaacatatga tgctggaatg 3241 gatatttggg gaggagaaaa cctagaaatt tcctttagga tttggcagtg tggaggaact 3301 ttggaaattg ttacatgctc acatgttgga catgtgtttc ggaaagctac accttacacg 3361 tttccaggag gcacagggca gattatcaat aaaaataaca gacgacttgc agaagtgtgg 3421 atggatgaat tcaagaattt cttctatata atttctccag gtgttacaaa ggtagattat 3481 ggagatatat cgtcaagagt tggtctaaga cacaaactac aatgcaaacc tttttcctgg 3541 tacctagaga atatatatcc tgattctcaa attccacgtc actatttctc attgggagag 3601 atacgaaatg tggaaacgaa tcagtgtcta gataacatgg ctagaaaaga gaatgaaaaa 3661 gttggaattt ttaattgcca tggtatgggg ggtaatcagg ttttctctta tactgccaac 3721 aaagaaatta gaacagatga cctttgcttg gatgtttcca aacttaatgg cccagttaca 3781 atgctcaaat gccaccacct aaaaggcaac caactctggg agtatgaccc agtgaaatta 3841 accctgcagc atgtgaacag taatcagtgc ctggataaag ccacagaaga ggatagccag 3901 gtgcccagca ttagagactg caatggaagt cggtcccagc agtggcttct tcgaaacgtc 3961 accctgccag aaatattcgg atccgactac aaagaccatg acggtgatta taaagatcat 4021 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc 4081 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 4141 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 4201 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 4261 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 4321 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 4381 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 4441 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 4501 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 4561 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 4621 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 4681 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 4741 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4801 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4861 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4921 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4981 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 5041 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 5101 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 5161 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 5221 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 5281 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 5341 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5401 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5461 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5521 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5581 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 5641 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5701 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5761 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 5821 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5881 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5941 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 6001 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 6061 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 6121 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6181 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6241 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6301 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6481 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6541 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6601 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6661 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6721 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6781 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6841 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6901 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7021 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7081 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7141 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7201 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7261 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7321 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7381 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7441 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7501 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7561 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7621 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7681 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7741 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7801 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7861 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7921 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7981 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8041 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8101 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8161 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8221 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8281 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8341 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8401 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8461 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8521 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8581 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8641 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8701 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8761 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8821 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8881 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8941 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9001 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9061 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9121 gctg //