LOCUS V044554 9117 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044554 VERSION V044554 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 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2297..3295 /label="Cd226(NM_178687)" /note="Cd226(NM_178687)" /gene="Cd226" CDS 3326..4045 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" promoter 4087..4298 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4311..4579 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4606..5205 /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 5212..5800 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5874..6107 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6179..6300 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6319..6454 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6492..6508) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6516..6532 /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(6540..6570) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6585..6606 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6894..7482) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7653..8513) /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(8514..8618) /gene="bla" /label="AmpR promoter" primer_bind 9092..9108 /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 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagcgaattc gccaccatgg cttatgttac ttggcttttg gctattcttc atgtgcacaa 2341 agcactgtgt gaagagacat tgtgggacac aacagttcgg ctttctgaga ctatgactct 2401 ggaatgtgta tatccattga cgcataactt aacccaggtg gagtggacca agaacactgg 2461 cacaaagaca gtgagcatag cagtttacaa ccctaaccat aatatgcata tagaatctaa 2521 ctacctccat agagtacact tcctaaactc aacagtgggg ttccgcaaca tgagcctttc 2581 cttttacaat gcctcagaag cagacattgg catctactcc tgcttgtttc atgctttccc 2641 aaatggacct tgggaaaaga agataaaagt agtctggtca gatagttttg agatagcagc 2701 accctcggat agctacctgt ctgcagaacc tggacaagat gtcacactca cttgccagct 2761 tccaaggact tggccagtgc aacaagtcat atgggaaaaa gtccagcccc atcaggtaga 2821 catcttagct tcctgtaacc tatctcaaga gacaagatac acttcaaagt acctaagaca 2881 aacaaggagc aactgtagcc aggggagcat gaagagcatc ctcatcattc caaatgccat 2941 ggccgctgac tcaggacttt acagatgtcg ctcagaggcc attacaggaa aaaacaagtc 3001 ctttgtcata aggctgatca taactgatgg tggaaccaat aaacatttta tccttcccat 3061 cgttggaggg ttagtttcac tgttacttgt catcctaatt atcatcattt tcattttata 3121 taacaggaag agacggagac aggtgagaat tccacttaaa gagcccaggg ataaacagag 3181 taaggtagcc accaactgca gaagtcctac ttctcccatc cagtctacag atgatgaaaa 3241 agaggacatt tatgtaaact atccaacttt ctctcgaaga ccaaaaccaa gactcggagg 3301 agggggatcc ggaggagggg gaagcatggt gagcaagggc gaggagctgt tcaccggggt 3361 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 3421 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg 3481 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt 3541 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 3601 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 3661 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 3721 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 3781 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat 3841 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 3901 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc 3961 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 4021 cggcatggac gagctgtaca agtaagcggc cgcgaaggat ctgcgatcgc tccggtgccc 4081 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca 4141 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact 4201 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga 4261 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca 4321 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg 4381 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa 4441 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc 4501 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg 4561 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac cgagtacaag 4621 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc 4681 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg 4741 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 4801 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 4861 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 4921 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 4981 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5041 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5101 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5161 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga caatcaacct 5221 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5281 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5341 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5401 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5461 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5521 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5581 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 5641 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5701 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5761 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 5821 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 5881 gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt 5941 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6001 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6061 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6121 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6181 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6241 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 6301 tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc 6361 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta 6421 ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat 6481 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 6541 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 6601 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 6661 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 6721 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 6781 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 6841 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 6901 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 6961 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 7021 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 7081 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 7141 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 7201 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 7261 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 7321 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 7381 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 7441 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 7501 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 7561 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 7621 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 7681 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 7741 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 7801 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 7861 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 7921 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 7981 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 8041 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 8101 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 8161 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 8221 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 8281 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 8341 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 8401 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 8461 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 8521 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 8581 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 8641 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg 8701 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc 8761 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc 8821 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt 8881 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac 8941 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg 9001 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg 9061 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg //