LOCUS V020260 9058 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020260 VERSION V020260 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 1960..2171 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2184..2452 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 2492..2501 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2501..2566 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2582..4465 /label="Jph4(NM_177049)" /note="Jph4(NM_177049)" /gene="Jph4" CDS 4481..4534 /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 4541..5140 /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 5153..5741 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5815..6048 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6120..6241 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6260..6395 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6433..6449) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6457..6473 /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(6481..6511) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6526..6547 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6835..7423) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7594..8454) /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(8455..8559) /gene="bla" /label="AmpR promoter" primer_bind 9033..9049 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca 1981 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc 2041 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg 2101 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg 2161 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta 2221 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct 2281 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc 2341 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc 2401 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt 2461 gaccggcgcc tactctagag ctagcgaatt cgccaccatg gactacaaag accatgacgg 2521 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagcttg gtaccctcga 2581 gatgtccccc gggggcaagt tcgactttga cgacgggggc tgctacgtgg ggggctggga 2641 ggcggggcgg gcacatggct acggcgtgtg caccgggccc ggcgcccagg gagagtacag 2701 cggctgctgg gctcacggct tcgagtcgct gggtgtcttc acggggcccg gcggacacag 2761 ttaccagggc cactggcagc agggcaagcg cgaagggctg ggcgtggagc gcaagagccg 2821 ctggacctac cgcggcgagt ggctgggcgg gctgaagggg cgcagcggcg tatgggagag 2881 cgtgtccggc ctgcgctacg ccgggctctg gaaggacggc ttccaggacg gctacggcac 2941 cgagacctac tcggatggag gcacttacca aggccagtgg caagccggga aacgccacgg 3001 ctacggggtg cgccaaagtg taccctacca ccaggcggcg cttctgcgct caccccgccg 3061 cacctccctg gactccggcc acagcgaccc cccgacgccg cctccacccc tacccctacc 3121 aggagatgaa ggaggcagcc cagcctccgg ctcccgaggc ggcttcgtgc tggcggggcc 3181 aggggacgcc gacggggcgt cttcccgtaa gcgcacccca gcagcaggtg gattcttccg 3241 ccgttcactg ctgctcagcg ggctccgggc tggtgggcgc cgcagctccc tgggcagcaa 3301 gaggggatct ctacgcagcg aggtgagcag cgaagtgggc agtacaggac ccccgggctc 3361 cgaggccagc gggcccccaa tcccagctcc acctgccctc atcgagggct cagccactga 3421 ggtgtacgcc ggtgagtggc gtgcggaccg gcgcagtggc tatggagtga gccagagatc 3481 caacgggctg cgctacgaag gcgagtggct gggcaaccga cgccacggat acgggcgcac 3541 cacccggccc gatggctccc gtgaggaggg caagtacaag cgtaaccggc tggtacacgg 3601 gggacgtgtc cgcagcctcc tgcctctggc ccttagacgg ggcaaagtca aggagaaagt 3661 ggacagggct gtcgaaggtg cccgtcgagc tgtgagtgct gcccgccagc gccaggaaat 3721 tgcggctgcc agggcagcag acgccctcct aaaagcagtg gcagccagca gcgtcgcgga 3781 gaaggctgtg gaggctgcgc ggatggccaa gctgatagcc caggatctac aacccatgtt 3841 agaggcccca ggccgcagac ccagacagga ctcaggaggt tctgacacag agcctttgga 3901 tgaggacagc cctggggtat acgagaatgg actgaccccc tcagagggct ctccggaact 3961 acccagcagc cccgcctcct cccaccaacc ttggcgagcc cctgcctgcc ggagtccact 4021 gcctcctgga gggaactggg gccccttctc cagccctaaa gcttggcccg aggagtgggg 4081 gggtcctggt gagcaggcag aggaactagc tggctatgag gctgaggatg aggctggggt 4141 gcagggtcca ggaccaagag atggttcccc acttctcgga ggctgcagtg acagttcagg 4201 aagtcttcga gaggaggagg gggaagacga agagtccttg ccccaactga gggccccagg 4261 aggctcagag tctgagcctg tcaccacgcc agttttgagg ggcctgtctt caaggggtcc 4321 tgatgctggg tgcctgacag aagaggttga ggagcctgct cccaccgaga ggcctgccca 4381 gccgggagct gccaaccccc tggtggtggg agccgtggcc ttgctggacc tcagcctggc 4441 cttcctgttc tcccagctcc tcaccggatc cgcggccgct gagggcagag gaagtcttct 4501 aacatgcggt gacgtggagg agaatcccgg cccttccggg atgaccgagt acaagcccac 4561 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt 4621 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac 4681 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc 4741 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt 4801 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca 4861 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac 4921 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg 4981 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa 5041 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg 5101 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga atctaggtcg acaatcaacc 5161 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 5221 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 5281 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 5341 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 5401 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 5461 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 5521 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt 5581 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 5641 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 5701 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa 5761 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag 5821 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt 5881 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 5941 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 6001 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat 6061 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga 6121 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 6181 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 6241 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc 6301 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct 6361 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa 6421 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6481 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6541 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6601 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6661 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6721 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6781 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6841 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6901 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 6961 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7021 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7081 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7141 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7201 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7261 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7321 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7381 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7441 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7501 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7561 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7621 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7681 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7741 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 7801 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7861 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 7921 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 7981 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8041 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8101 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8161 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8221 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8281 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8341 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8401 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8461 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8521 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8581 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8641 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8701 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 8761 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 8821 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 8881 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 8941 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9001 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //