LOCUS V030221 9658 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030221 VERSION V030221 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..3684 /label="SLC18B1(NM_052831)" /note="SLC18B1(NM_052831)" /gene="SLC18B1" CDS 3691..3756 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3800..4011 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4024..4292 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4321..4330 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4351..5013 /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 5083..5136 /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 5137..5736 /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 5737..6325 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6399..6632 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6704..6825 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6865..7000 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7033..7049) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7057..7073 /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(7081..7111) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7126..7147 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7435..8023) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8194..9054) /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(9055..9159) /gene="bla" /label="AmpR promoter" primer_bind 9633..9649 /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 gccaccatgg aggcgctggg tgacctggag 2341 ggaccacgcg caccaggagg tgatgatcct gcaggaagtg caggagagac ccccgggtgg 2401 ctttcgagag aacaggtttt tgtactgata tcggcagctt cggtgaactt aggttccatg 2461 atgtgctatt ctatacttgg accgtttttc cccaaagagg ctgaaaagaa gggagccagc 2521 aatacaatta tcggtatgat ctttggatgt tttgctttgt tcgagttgct ggcatccttg 2581 gtatttggaa actatcttgt acatattgga gcaaaattta tgtttgtagc aggaatgttt 2641 gtctcaggag gagttacaat tctctttggt gtattggacc gagttccaga tgggccagta 2701 tttattgcta tgtgttttct agtgagagta atggatgcag ttagctttgc tgcagcaatg 2761 actgcatctt cttctatcct ggcaaaggct tttccaaata acgtggctac ggtattggga 2821 agtcttgaga ctttttctgg actggggcta atactaggtc ctcctgtagg tggctttttg 2881 tatcaatcct ttggctatga agtgcctttt attgttctgg gatgcgtcgt tttgctgatg 2941 gtaccactca atatgtatat tttacccaat tacgagtctg atccaggtga acactcattc 3001 tggaaactga tcgctttacc caaagttggc cttatagcct tcgtcatcaa ctcactcagc 3061 tcgtgttttg gcttcctcga tcctactctg tctctctttg ttttggagaa gttcaattta 3121 ccagctggat atgtgggact agtattcctg ggtatggcac tgtcctatgc catctcttca 3181 ccactatttg gtctcctaag tgataaaagg ccacctctaa ggaaatggct tctggtgttt 3241 ggcaacttaa tcacagccgg gtgctacatg ctcttagggc ctgtcccaat cttgcatatt 3301 aaaagtcagc tctggctgct ggtgctgata ttagttgtaa gtggcctctc tgctggaatg 3361 agtataattc caactttccc ggaaattctc agttgtgcac atgaaaatgg gtttgaagag 3421 ggattaagta cattgggact tgtatcaggt ctttttagtg caatgtggtc aattggtgct 3481 tttatgggac caacgctggg tggatttctg tatgagaaaa ttggttttga atgggcagca 3541 gctatacaag gtctatgggc tctgataagt ggattagcca tgggcttgtt ttatctactg 3601 gagtattcaa ggagaaaaag gtctaaatct caaaacatcc tcagcacaga ggaggaacga 3661 actactctct tgcctaatga aaccggatcc gactacaaag accatgacgg tgattataaa 3721 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3781 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3841 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3901 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3961 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 4021 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 4081 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4141 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4201 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4261 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 4321 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 4381 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 4441 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 4501 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4561 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4621 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4681 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4741 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4801 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4861 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4921 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4981 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 5041 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 5101 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 5161 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 5221 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 5281 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 5341 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 5401 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 5461 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 5521 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5581 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5641 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5701 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5761 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5821 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5881 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5941 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 6001 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 6061 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 6121 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 6181 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 6241 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 6301 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 6361 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 6421 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 6481 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6541 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6601 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6661 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6721 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6781 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6841 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6901 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6961 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 7021 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 7081 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 7141 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 7201 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 7261 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 7321 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 7381 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 7441 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7501 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7561 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7621 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7681 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7741 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7801 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7861 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7921 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7981 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 8041 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 8101 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 8161 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 8221 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 8281 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 8341 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 8401 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 8461 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8521 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8581 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8641 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8701 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8761 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8821 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8881 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8941 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 9001 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 9061 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 9121 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 9181 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 9241 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 9301 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 9361 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 9421 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9481 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9541 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9601 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //