LOCUS V044759 9548 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044759 VERSION V044759 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2290..3690 /label="Smad2(NM_010754)" /note="Smad2(NM_010754)" /gene="Smad2" CDS 3727..4443 /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" CDS 4450..4467 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" promoter 4518..4729 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4742..5010 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5037..5636 /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 5643..6231 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6305..6538 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6610..6731 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6750..6885 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6923..6939) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6947..6963 /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(6971..7001) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7016..7037 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7325..7913) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8084..8944) /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(8945..9049) /gene="bla" /label="AmpR promoter" primer_bind 9523..9539 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagccacca tgtcgtccat cttgccattc actccgccag tggtgaagag acttctggga 2341 tggaaaaaat cagccggtgg gtctggagga gcaggtggtg gagagcagaa tggacaggaa 2401 gaaaagtggt gtgaaaaagc agtgaaaagt ctggtgaaaa agctaaagaa aacaggacgg 2461 ttagatgagc ttgagaaagc catcaccact cagaattgca atactaaatg tgtcaccata 2521 ccaagcactt gctctgaaat ttggggactg agtacagcaa atacggtaga tcagtgggac 2581 acaacaggcc tttacagctt ctctgaacaa accaggtctc ttgatggccg tcttcaggtt 2641 tcacaccgga aagggttgcc acatgttata tattgccggc tctggcgctg gccggacctt 2701 cacagtcatc atgagctcaa ggcaatcgaa aactgcgaat atgcttttaa tctgaaaaaa 2761 gatgaagtgt gtgtaaatcc gtaccactac cagagagttg agaccccagt cttgcctcca 2821 gtcttagtgc ctcggcacac ggagattcta acagaactgc cgcccctgga tgactacacc 2881 cactccattc cagaaaacac aaatttccca gcaggaattg agccacagag taattacatc 2941 ccagaaacac caccacctgg atatatcagt gaagatggag aaacaagtga ccaacagttg 3001 aaccaaagta tggacacagg ctctccggct gaactgtctc ctactactct ctctcctgtt 3061 aatcacagct tggatttgca gccagttact tactcggaac ctgcattctg gtgttcaatc 3121 gcatactatg aactaaacca gagggttgga gagaccttcc atgcgtcaca gccctcgctc 3181 actgtagacg gcttcacaga cccatcaaac tcggagaggt tctgcttagg cttgctctcc 3241 aacgttaacc gaaatgccac tgtagaaatg acaagaagac atataggaag gggagtgcgc 3301 ttgtattaca taggtgggga agtgtttgct gagtgcctaa gtgatagtgc aatctttgtg 3361 cagagcccca actgtaacca gagatacggc tggcaccctg caacagtgtg taagatccca 3421 ccaggctgta acctgaagat cttcaacaac caagaatttg ctgctcttct ggctcagtct 3481 gtcaaccagg gttttgaagc cgtttatcag ctaacccgaa tgtgcaccat aagaatgagt 3541 tttgtgaagg gctggggagc agaatatcgg aggcagacag taacaagtac tccttgctgg 3601 attgaacttc atctgaatgg ccctctgcag tggctggaca aagtattaac tcagatggga 3661 tccccttcag tgcgatgctc aagcatgtcg gaattcggag gtggcggaag cggcggtgga 3721 ggtagtatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag 3781 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc 3841 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg 3901 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac 3961 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc 4021 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac 4081 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg 4141 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag 4201 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag 4261 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac 4321 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac 4381 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac 4441 aagctcgagc accaccacca ccaccactga ggatccgcgg ccgcgaagga tctgcgatcg 4501 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg 4561 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga 4621 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt 4681 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg 4741 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg 4801 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta 4861 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc 4921 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt 4981 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga 5041 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca 5101 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc 5161 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 5221 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 5281 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 5341 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 5401 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 5461 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 5521 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 5581 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg 5641 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 5701 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 5761 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 5821 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 5881 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 5941 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 6001 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc 6061 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 6121 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 6181 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct 6241 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 6301 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg 6361 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6421 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6481 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 6541 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 6601 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 6661 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 6721 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 6781 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 6841 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 6901 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 6961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 7021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 7081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 7141 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 7201 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 7261 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 7321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 7381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 7441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 7501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 7561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 7621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 7681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 7741 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 7801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 7861 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 7921 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 7981 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 8041 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 8101 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 8161 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 8221 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 8281 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 8341 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 8401 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8461 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 8521 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 8581 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 8641 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 8701 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 8761 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 8821 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 8881 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 8941 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 9001 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 9061 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 9121 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 9181 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 9241 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 9301 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 9361 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 9421 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 9481 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 9541 ccaagctg //