LOCUS V040947 9848 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040947 VERSION V040947 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)" CDS 2498..4024 /label="SOX9(NM_000346)" /note="SOX9(NM_000346)" /gene="SOX9" CDS 4031..4087 /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 porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 4088..4807 /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 4817..5316 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5337..5936 /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 5943..6531 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6605..6838 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6910..7031 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7050..7185 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7223..7239) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7247..7263 /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(7271..7301) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7316..7337 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7625..8213) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8384..9244) /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(9245..9349) /gene="bla" /label="AmpR promoter" primer_bind 9823..9839 /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 aatctcctgg accccttcat 2521 gaagatgacc gacgagcagg agaagggcct gtccggcgcc cccagcccca ccatgtccga 2581 ggactccgcg ggctcgccct gcccgtcggg ctccggctcg gacaccgaga acacgcggcc 2641 ccaggagaac acgttcccca agggcgagcc cgatctgaag aaggagagcg aggaggacaa 2701 gttccccgtg tgcatccgcg aggcggtcag ccaggtgctc aaaggctacg actggacgct 2761 ggtgcccatg ccggtgcgcg tcaacggctc cagcaagaac aagccgcacg tcaagcggcc 2821 catgaacgcc ttcatggtgt gggcgcaggc ggcgcgcagg aagctcgcgg accagtaccc 2881 gcacttgcac aacgccgagc tcagcaagac gctgggcaag ctctggagac ttctgaacga 2941 gagcgagaag cggcccttcg tggaggaggc ggagcggctg cgcgtgcagc acaagaagga 3001 ccacccggat tacaagtacc agccgcggcg gaggaagtcg gtgaagaacg ggcaggcgga 3061 ggcagaggag gccacggagc agacgcacat ctcccccaac gccatcttca aggcgctgca 3121 ggccgactcg ccacactcct cctccggcat gagcgaggtg cactcccccg gcgagcactc 3181 ggggcaatcc cagggcccac cgaccccacc caccaccccc aaaaccgacg tgcagccggg 3241 caaggctgac ctgaagcgag aggggcgccc cttgccagag gggggcagac agccccctat 3301 cgacttccgc gacgtggaca tcggcgagct gagcagcgac gtcatctcca acatcgagac 3361 cttcgatgtc aacgagtttg accagtacct gccgcccaac ggccacccgg gggtgccggc 3421 cacgcacggc caggtcacct acacgggcag ctacggcatc agcagcaccg cggccacccc 3481 ggcgagcgcg ggccacgtgt ggatgtccaa gcagcaggcg ccgccgccac ccccgcagca 3541 gcccccacag gccccgccgg ccccgcaggc gcccccgcag ccgcaggcgg cgcccccaca 3601 gcagccggcg gcacccccgc agcagccaca ggcgcacacg ctgaccacgc tgagcagcga 3661 gccgggccag tcccagcgaa cgcacatcaa gacggagcag ctgagcccca gccactacag 3721 cgagcagcag cagcactcgc cccaacagat cgcctacagc cccttcaacc tcccacacta 3781 cagcccctcc tacccgccca tcacccgctc acagtacgac tacaccgacc accagaactc 3841 cagctcctac tacagccacg cggcaggcca gggcaccggc ctctactcca ccttcaccta 3901 catgaacccc gctcagcgcc ccatgtacac ccccatcgcc gacacctctg gggtcccttc 3961 catcccgcag acccacagcc cccagcactg ggaacaaccc gtctacacac agctcactcg 4021 acctggatcc gccactaact tctccctgtt gaaacaagca ggggatgtcg aagagaatcc 4081 cgggccaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga 4141 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc 4201 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg 4261 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca 4321 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac 4381 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga 4441 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct 4501 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca 4561 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca 4621 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga 4681 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca 4741 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta 4801 caagtaggcg gccgctgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct 4861 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc 4921 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct 4981 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga 5041 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg 5101 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct 5161 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg 5221 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc 5281 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag cttaccatga 5341 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca 5401 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc 5461 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 5521 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 5581 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 5641 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 5701 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 5761 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 5821 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 5881 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg 5941 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 6001 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 6061 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 6121 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 6181 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 6241 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 6301 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc 6361 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 6421 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 6481 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct 6541 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 6601 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg 6661 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6721 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6781 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 6841 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 6901 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 6961 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 7021 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 7081 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 7141 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 7201 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 7261 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 7321 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 7381 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 7441 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 7501 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 7561 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 7621 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 7681 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 7741 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 7801 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 7861 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 7921 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 7981 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 8041 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 8101 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 8161 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 8221 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 8281 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 8341 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 8401 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 8461 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 8521 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 8581 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 8641 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 8701 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8761 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 8821 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 8881 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 8941 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 9001 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 9061 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 9121 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 9181 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 9241 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 9301 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 9361 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 9421 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 9481 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 9541 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 9601 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 9661 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 9721 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 9781 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 9841 ccaagctg //