LOCUS V037944 9766 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037944 VERSION V037944 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..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2317..3792 /label="SLC39A14(NM_001128431)" /note="SLC39A14(NM_001128431)" /gene="SLC39A14" CDS 3799..3864 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3908..4119 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4132..4400 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4429..4438 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4459..5121 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 5191..5244 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 5245..5844 /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 5845..6433 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6507..6740 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6812..6933 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6973..7108 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7141..7157) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7165..7181 /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(7189..7219) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7234..7255 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7543..8131) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8302..9162) /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(9163..9267) /gene="bla" /label="AmpR promoter" primer_bind 9741..9757 /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 attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc ctgatacgaa ctcggaattc gccaccatga agctgctgct gctgcacccg 2341 gccttccaga gctgcctcct gctgaccctg cttggcttat ggagaaccac ccctgaggct 2401 cacgcttcat ccctgggtgc accagctatc agcgctgcct ccttcctgca ggatctaata 2461 catcggtatg gcgagggtga cagcctcact ctgcagcagc tgaaggccct actcaaccac 2521 ctggatgtgg gagtgggccg gggtaatgtc acccagcacg tgcaaggaca caggaacctc 2581 tccacgtgct ttagttctgg agacctcttc actgcccaca atttcagcga gcagtcgcgg 2641 attgggagca gcgagctcca ggagttctgc cccaccatcc tccagcagct ggattcccgg 2701 gcctgcacct cggagaacca ggaaaacgag gagaatgagc agacggagga ggggcggcca 2761 agcgctgttg aagtgtgggg atacggtctc ctctgtgtga ccgtcatctc cctctgctcc 2821 ctcctggggg ccagcgtggt gcccttcatg aagaagacct tttacaagag gctgctgctc 2881 tacttcatag ctctggcgat tggaaccctc tactccaacg ccctcttcca gctcatcccg 2941 gaggcatttg gtttcaaccc tctggaagat tattatgtct ccaagtctgc agtggtgttt 3001 gggggctttt atcttttctt tttcacagag aagatcttga agattcttct taagcagaaa 3061 aatgagcatc atcatggaca cagccattat gcctctgagt cgcttccctc caagaaggac 3121 caggaggagg gggtgatgga gaagctgcag aacggggacc tggaccacat gattcctcag 3181 cactgcagca gtgagctgga cggcaaggcg cccatggtgg acgagaaggt cattgtgggc 3241 tcgctctctg tgcaggacct gcaggcttcc cagagtgctt gctactggct gaaaggtgtc 3301 cgctactctg atatcggcac tctggcctgg atgatcactc tgagcgacgg cctccataat 3361 ttcatcgatg gcctggccat cggtgcttcc ttcactgtgt cagttttcca aggcatcagc 3421 acctcggtgg ccatcctctg tgaggagttc ccacatgagc taggagactt tgtcatcctg 3481 ctcaacgctg ggatgagcat ccaacaagct ctcttcttca acttcctttc tgcctgctgc 3541 tgctacctgg gtctggcctt tggcatcctg gccggcagcc acttctctgc caactggatt 3601 tttgcgctag ctggaggaat gttcttgtat atttctctgg ctgatatgtt ccctgagatg 3661 aatgaggtct gtcaagagga tgaaaggaag ggcagcatct tgattccatt tatcatccag 3721 aacctgggcc tcctgactgg attcaccatc atggtggtcc tcaccatgta ttcaggacag 3781 atccagattg ggggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3841 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3901 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3961 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 4021 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 4081 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 4141 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 4201 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 4261 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 4321 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4381 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 4441 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 4501 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4561 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4621 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4681 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4741 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 4801 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 4861 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 4921 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 4981 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 5041 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 5101 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 5161 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 5221 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 5281 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5341 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5401 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5461 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5521 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5581 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5641 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5701 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5761 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5821 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 5881 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 5941 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 6001 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 6061 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 6121 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 6181 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 6241 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 6301 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 6361 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6421 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6481 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6541 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6601 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6661 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6721 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 6781 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 6841 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 6901 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 6961 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 7021 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 7081 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 7141 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 7201 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 7261 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7321 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7381 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7441 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7501 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7561 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7621 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7681 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7741 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7801 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7861 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7921 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7981 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 8041 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 8101 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 8161 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 8221 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 8281 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8341 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8401 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8461 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8521 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8581 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8641 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8701 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8761 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8821 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8881 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8941 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 9001 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 9061 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 9121 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 9181 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 9241 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 9301 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 9361 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9421 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9481 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9541 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9601 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9661 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9721 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //