LOCUS V027657 10116 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027657 VERSION V027657 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 1494..1538 /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 1801..1916 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 1974..2277 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2278..2481 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2588..4186 /label="Phgdh(NM_016966)" /note="Phgdh(NM_016966)" /gene="Phgdh" CDS 4211..4282 /codon_start=1 /product="three tandem FLAG® epitope tags" /transl_table=1 /translation="DYKDDDDKDYKDDDDKDYKDDDDK" /label="3xFLAG" promoter 4336..4547 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4560..4828 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4856..5548 /codon_start=1 /note="mammalian codon-optimized" /product="Zoanthus green fluorescent protein" /transl_table=1 /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH AIASGSALP" /label="ZsGreen1" CDS 5558..5611 /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 5612..6211 /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 6212..6799 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6873..7106 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7178..7299 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7318..7453 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7491..7507) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7515..7531 /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(7539..7569) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7584..7605 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7893..8481) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8652..9512) /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(9513..9617) /gene="bla" /label="AmpR promoter" primer_bind 10091..10107 /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 gggcgcagcg 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 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca 1441 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg 1501 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa 1561 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa 1621 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt 1681 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag 1741 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgcc 1801 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata 1861 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg 1921 gtttattaca gggacagcag agatccagtt tatcgataag cttgggagtt ccgcgttaca 1981 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 2041 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 2101 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 2161 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 2221 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 2281 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 2341 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2401 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2461 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca 2521 cgctgttttg acctccatag aagacaccga ctctactaga ggatctattt ccggtgaatt 2581 cgccaccatg gccttcgcaa atctgcgcaa agtactcatc agtgacagcc tggacccctg 2641 ctgccggaag atcctgcaag atggagggct gcaggtggtg gagaagcaga acttgagcaa 2701 ggaggagctg atagctgaac tccaggactg tgaaggcctc attgtccggt ctgctactaa 2761 ggtcactgct gatgtcatca atgcagcaga gaagctccag gtggtgggca gggctggcac 2821 aggtgtggac aatgtggatc tggaggctgc cacgaggaag ggcatcctag tcatgaacac 2881 ccccaacggg aacagcctca gtgctgcgga gctcacctgt gggatgatca tgtgcctggc 2941 caggcagatt ccccaggcaa cagcttcgat gaaagatggc aaatgggacc ggaagaagtt 3001 catggggaca gaactgaacg ggaagacact gggaattctt ggcctcggca gaattggaag 3061 agaggtggcc acccgaatgc aatcctttgg aatgaagact gtaggctatg accccatcat 3121 ctctcctgaa gtcgccgcct cctttggtgt tcagcagctg ccgctggagg agatctggcc 3181 tctctgtgac tttataactg tccatacccc actcctgccc tctaccacag gcttgctgaa 3241 tgacagcacc tttgctcagt gcaagaaagg tgtgcgagta gtgaactgtg ctcgaggagg 3301 cattgtggat gaaggtgccc tgctgcgtgc cctgcagtct ggtcagtgtg ctggggctgc 3361 actggatgtg tttacagaag agccaccacg ggaccgggcc ttagtggacc acgagaatgt 3421 catcagctgt ccccacctgg gtgccagcac caaggaagcc cagagccgct gtggggagga 3481 aatcgcagtc cagtttgtgg acatggtgaa ggggaaatct ctaacaggcg ttgtgaacgc 3541 ccaggccctc accagtgcct tctctccaca caccaagcct tggattggtc tggcagaagc 3601 aatgggcacg ctgatgcacg cctgggctgg ctcccctaaa gggaccatcc aggtggttac 3661 acaaggaaca tctctgaaga atgctgggac ctgcctgagc cctgcagtca ttgtcggcct 3721 tctgagagaa gcatctaagc aggcagacgt gaacttggtg aacgctaagc tactggtgaa 3781 agaggctggc ctcaatgtca ccacctccca caaccctggg gttccagggg agcagggtag 3841 cggggaatgc ctcctgactg tggccctggc aggtgctccc taccaagccg tgggcttggt 3901 ccagggcacc acaccaatgc tgcagatgct caacggagct gtcttcagac cagaggtgcc 3961 actacgcagg ggccaacccc tgctcgtatt ccgggctcag ccctccgacc ctgggatgct 4021 gcccactatg attggcctcc tggcagaggc gggtgtacag ctgctgtcct accaaacctc 4081 catggtgtct gacggagagc cctggcacgt catgggcctg tcctccctgc tgcccagcct 4141 ggaaacatgg aagcagcatg tattggaggc tttccagttc tgcttcggat cctcggtacc 4201 aagcttaagt gactacaagg atgacgatga caaggattac aaagacgacg atgataagga 4261 ctataaggat gatgacgaca aataaagatc catcgatact agtaaggatc tgcgatcgct 4321 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 4381 gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 4441 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 4501 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag 4561 gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt 4621 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg 4681 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta 4741 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt 4801 cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctact ctagcatggc 4861 ccagtccaag cacggcctga ccaaggagat gaccatgaag taccgcatgg agggctgcgt 4921 ggacggccac aagttcgtga tcaccggcga gggcatcggc taccccttca agggcaagca 4981 ggccatcaac ctgtgcgtgg tggagggcgg ccccttgccc ttcgccgagg acatcttgtc 5041 cgccgccttc atgtacggca accgcgtgtt caccgagtac ccccaggaca tcgtcgacta 5101 cttcaagaac tcctgccccg ccggctacac ctgggaccgc tccttcctgt tcgaggacgg 5161 cgccgtgtgc atctgcaacg ccgacatcac cgtgagcgtg gaggagaact gcatgtacca 5221 cgagtccaag ttctacggcg tgaacttccc cgccgacggc cccgtgatga agaagatgac 5281 cgacaactgg gagccctcct gcgagaagat catccccgtg cccaagcagg gcatcttgaa 5341 gggcgacgtg agcatgtacc tgctgctgaa ggacggtggc cgcttgcgct gccagttcga 5401 caccgtgtac aaggccaagt ccgtgccccg caagatgccc gactggcact tcatccagca 5461 caagctgacc cgcgaggacc gcagcgacgc caagaaccag aagtggcacc tgaccgagca 5521 cgccatcgcc tccggctccg ccttgcccgc ggccgctgag ggcagaggaa gtcttctaac 5581 atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct 5641 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta 5701 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca 5761 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg 5821 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga 5881 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga 5941 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt 6001 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc 6061 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt 6121 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac 6181 ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtga 6241 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 6301 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 6361 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 6421 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 6481 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 6541 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 6601 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg 6661 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 6721 gctgccggct ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga gtcggatctc 6781 cctttggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga 6841 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa 6901 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga 6961 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct 7021 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt 7081 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt 7141 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa 7201 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 7261 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc 7321 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 7381 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 7441 ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg 7501 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 7561 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 7621 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 7681 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 7741 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 7801 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 7861 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 7921 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 7981 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 8041 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 8101 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 8161 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 8221 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 8281 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 8341 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 8401 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 8461 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 8521 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 8581 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 8641 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 8701 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 8761 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 8821 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 8881 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 8941 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 9001 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 9061 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 9121 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 9181 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 9241 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 9301 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 9361 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 9421 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 9481 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 9541 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 9601 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt 9661 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt 9721 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt 9781 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg 9841 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg 9901 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca 9961 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag 10021 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag 10081 tcacgacgtt gtaaaacgac ggccagtgcc aagctg //