LOCUS V042339 9418 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042339 VERSION V042339 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..3444 /label="Fmod(NM_021355)" /note="Fmod(NM_021355)" /gene="Fmod" CDS 3451..3516 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3560..3771 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3784..4052 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4081..4090 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4111..4773 /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 4843..4896 /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 4897..5496 /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 5497..6085 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6159..6392 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6464..6585 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6625..6760 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6793..6809) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6817..6833 /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(6841..6871) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6886..6907 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7195..7783) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7954..8814) /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(8815..8919) /gene="bla" /label="AmpR promoter" primer_bind 9393..9409 /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 gccaccatgc agtgggcctc cgtcctgctg 2341 ctggctgggc tctgctcact ttcccagggc cagtatgatg aagactctca ctggtggatc 2401 caatacctcc gaaaccagca gtccacctac tacgacccct atgaccctta cccctatgag 2461 ccctctgagc cttaccccta tggagtggaa gaaggcccag cctatgccta tggtgcacca 2521 cctccaccag agccccgtga ttgtccccaa gaatgcgact gtccccccaa cttccccaca 2581 gccatgtact gtgacaaccg caacctcaag tacctgcctt ttgtgccctc ccgcatgaag 2641 tacgtctact tccagaacaa ccagatctct gccatccagg aaggtgtctt tgacaatgcc 2701 actgggctcc tctgggtcgc tctacatggc aaccagatta ccagtgacaa ggtaggcagg 2761 aaagtcttct ccaagctgag gcacttggag aggctgtact tggaccacaa caacctgacc 2821 cggatgcccg gcccgctgcc ccgatccctg agagagctcc acctggacca caaccagatc 2881 tcacgggtcc ccaacaatgc tttggagggc ctggagaacc tcacggcctt atatctccac 2941 cacaatgaga tccaggaagt ggggagttcc atgagaggcc tccggtccct gatcctacta 3001 gacctgagtt ataaccacct tcggagggta cccgacggtc tgccctcagc cctggagcag 3061 ctgtacctag aacacaacaa tgtctacacc gtccctgaca gctacttccg ggggtcaccc 3121 aagctgctgt acgtccggtt gtctcacaac agtctcacta acaacggcct tgctaccaac 3181 accttcaact ccagcagcct tcttgagcta gacttgtctt acaatcagct gcagaagatc 3241 cctcctgtca acaccaacct ggagaatctt tacctccagg gcaacaggat caatgagttc 3301 tccatcagca gcttctgcac ggtggtggac gtcatgaact tctccaagct gcaggtgcta 3361 cgcctggatg ggaacgagat caagcgcagc gcaatgccgg tggacgcccc actctgcctg 3421 cgcctcgcca acctcatcga gatcggatcc gactacaaag accatgacgg tgattataaa 3481 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3541 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3601 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3661 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3721 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3781 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3841 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3901 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3961 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4021 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 4081 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 4141 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 4201 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 4261 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4321 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4381 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4441 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4501 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4561 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4621 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4681 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 4741 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 4801 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 4861 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 4921 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 4981 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 5041 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 5101 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 5161 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 5221 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 5281 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5341 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5401 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5461 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5521 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5581 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5641 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5701 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5761 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5821 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5881 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 5941 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 6001 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 6061 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 6121 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 6181 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 6241 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6301 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6361 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6421 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6481 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6541 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6601 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6661 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6721 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 6781 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6841 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6901 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6961 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 7021 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 7081 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 7141 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 7201 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7261 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7321 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7381 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7441 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7501 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7561 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7621 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7681 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7741 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7801 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7861 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7921 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7981 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 8041 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 8101 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 8161 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 8221 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8281 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8341 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8401 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8461 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8521 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8581 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8641 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8701 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8761 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8821 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8881 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8941 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 9001 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 9061 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 9121 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 9181 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9241 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9301 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9361 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //