LOCUS V045037 9962 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045037 VERSION V045037 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers polyA_signal 216..350 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" promoter 398..502 /gene="bla" /label="AmpR promoter" CDS 503..1363 /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" rep_origin 1534..2119 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" protein_bind 2407..2428 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 2443..2473 /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 2481..2497 /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" primer_bind 2505..2521 /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" LTR 2650..3283 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 3490..4078 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" CDS complement(4092..4691) /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" promoter complement(4700..5029) /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS complement(5040..5786) /codon_start=1 /product="modified rtTA protein that binds tightly to promoters containing the tet operator in the presence of doxycycline" /transl_table=1 /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG*" /label="Tet-On® 3G" promoter complement(5805..6315) /note="human phosphoglycerate kinase 1 promoter" /label="hPGK promoter" promoter 6332..6699 /note="3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On® 3G, modified to eliminate binding sites for endogenous mammalian transcription factors" /label="TRE3GS promoter" CDS 6742..7461 /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" polyA_signal 7642..7776 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" misc_feature 7820..7935 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" CDS complement(8198..8242) /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 8427..8660 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" misc_feature 9157..9282 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" LTR 9329..9962 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" ORIGIN 1 actactaagt ttgtagtaca tatttaacaa atacaatttc tttaaaaact actaagtttg 61 tagtacatat ttaacaaata caatttcttt aaaatgaaaa taattcagag gaatcacagg 121 tttagagtaa atgaaaccac taggtaattg gcagtggtaa tagggtatgg ggtgggaagt 181 ttgggatgat tttggttagc ttgagttatc cagttgatcc agacatgata agatacattg 241 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt 301 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt gatctcccga 361 tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 421 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 481 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt 541 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga 601 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa 661 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct 721 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat 781 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga 841 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc 901 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat 961 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa 1021 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac 1081 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa 1141 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc 1201 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc 1261 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag 1321 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta 1381 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 1441 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 1501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 1561 ctgctgcttg caaacaaaaa aaccgctacc agcggtggtt tgtttgccgg atcaagagct 1621 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 1681 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 1741 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 1801 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 1861 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 1921 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 1981 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 2041 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 2101 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 2161 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 2221 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 2281 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 2341 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 2401 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 2461 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 2521 ccatgattac gccaagctct agctagaggt cgacggtaaa acgctagcaa tgtcaaggcc 2581 tctcactctc tgatattcat ttctttgcaa gttataaata ctgaataata agatgacatg 2641 aactactact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac 2701 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag 2761 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc 2821 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac 2881 tccccagtcc cgcccaggcc acgcctccct ggaaagtccc cagcggaaag tcccttgtag 2941 caagctcgat atcagcagtt cttgaagtac tccggatgca gctctcgggc cacgtgatga 3001 aatgctaggc ggctgtcaaa cctccactct aacacttctc tctccgggtc atccatccca 3061 tgcaggctca cagggtgtaa caagctggtg ttctctcctt tattggcctc ttctacctta 3121 tctggctcaa ctggtactag cttgtagcac catccaaagg tcagtggata tctgacccct 3181 ggccctggtg tgtagttctg ctaatcaggg aagtagcctt gtgtgtggta gatccacaga 3241 tcaaggatat cttgtcttcg ttgggagtga attagccctt ccagtcccct cttttctttt 3301 aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg tacctgaggt 3361 gtgactggaa aacccacctc ctcctcctct tgtgcttcta gccaggcaca atcagcattg 3421 gtagctgctg tattgctact tgtgattgct ccatgttttt ctaggtctcg actgcagaat 3481 taattccagg cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa 3541 gacgcggaag aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga 3601 gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga 3661 gcagccatgg aaaggacgtc agcttccccg acaacaccac ggaattgtca gtgcccaaca 3721 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag 3781 ggagggggaa agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag 3841 tgggggttgc gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca 3901 actcctcata aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac 3961 gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag 4021 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattgt 4081 tccagacgcg ttcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc 4141 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc 4201 gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc 4261 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg 4321 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg 4381 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc 4441 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac 4501 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc 4561 tcgatgtggc ggtccgggtc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg 4621 gcgagggtgc gtacggcccg ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg 4681 tactcggtca tggtcgcgtt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact 4741 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc 4801 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta 4861 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg 4921 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac 4981 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttcgcgcgtt 5041 tacccgggga gcatgtcaag gtcaaaatcg tcaagagcgt cagcaggcag catatcaagg 5101 tcaaagtcgt caagggcatc ggctgggagc atgtctaagt caaaatcgtc aagggcgtcg 5161 gtcggcccgc cgctttcgca ctttagctgt ttctccaggc cacatatgat tagttccagg 5221 ccgaaaagga aggcaggttc ggctccctgc cggtcgaaca gctcaattgc ttgtttcaga 5281 agtgggggca tagaatcggt ggtaggtgtc tctctttcct cttttgctac ttgatgctcc 5341 tgttcctcca atacgcagcc cagtgtaaag tggcccacgg cggacagagc gtacagtgcg 5401 ttctccaggg agaagccttg ctgacacagg aacgcgagct gattttccag ggtttcgtac 5461 tgtttctctg ttgggcgggt gccgagatgc actttagccc cgtcgcgatg tgagaggaga 5521 gcacagcggt atgacttggc gttgttccgc agaaagtctt gccatgactc gccttccagg 5581 gggcaggagt gggtatgatg cctgtccagc atctcgattg gcagggcatc gagcagggcc 5641 cgcttgttct tcacgtgcca gtacagggta ggctgctcaa ctcccagctt ttgagcgagt 5701 ttccttgtcg tcaggccttc gataccgact ccattgagta attccagagc agagtttatg 5761 actttgctct tgtccagtct agacatggta attcgatgat ccccctgggg agagaggtcg 5821 gtgattcggt caacgaggga gccgactgcc gacgtgcgct ccggaggctt gcagaatgcg 5881 gaacaccgcg cgggcaggaa cagggcccac actaccgccc cacaccccgc ctcccgcacc 5941 gccccttccc ggccgctgct ctcggcgcgc cctgctgagc agccgctatt ggccacagcc 6001 catcgcggtc ggcgcgctgc cattgctccc tggcgctgtc cgtctgcgag ggtactagtg 6061 agacgtgcgg cttccgtttg tcacgtccgg cacgccgcga accgcaagga accttcccga 6121 cttaggggcg gagcaggaag cgtcgccggg gggcccacaa gggtagcggc gaagatccgg 6181 gtgacgctgc gaacggacgt gaagaatgtg cgagacccag ggtcggcgcc gctgcgtttc 6241 ccggaaccac gcccagagca gccgcgtccc tgcgcaaacc cagggctgcc ttggaaaagg 6301 cgcaacccca accccgtgga attatcacct cgagtttact ccctatcagt gatagagaac 6361 gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca 6421 gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt 6481 tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac 6541 gtatatccag tttactccct atcagtgata gagaacgtat aagctttgct tatgtaaacc 6601 agggcgccta taaaagagtg ctgatttttt gagtaaactt caattccaca acacttttgt 6661 cttataccaa ctttccgtac cacttcctac cctcgtaaag aattcaccgg tggcggtgga 6721 ggaagcggcg gaggtggatc tatggtgagc aagggcgagg agctgttcac cggggtggtg 6781 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 6841 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 6901 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 6961 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 7021 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 7081 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 7141 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 7201 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 7261 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 7321 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 7381 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 7441 atggacgagc tgtacaagta gggatccaga ccacctcccc tgcgagctaa gctggacagc 7501 caatgacggg taagagagtg acatttttca ctaacctaag acaggagggc cgtcagagct 7561 actgcctaat ccaaagacgg gtaaaagtga taaaaatgta tcactccaac ctaagacagg 7621 cgcagcttcc gagggatttg agatccagac atgataagat acattgatga gtttggacaa 7681 accaaaacta gaatgcagtg aaaaaaatgc cttatttgtg aaatttgtga tgctattgcc 7741 ttatttgtaa ccattataag ctgcaataaa caagttaagt cgataaactg gatctctgct 7801 gtccctgtaa taaacccgaa aattttgaat ttttgtaatt tgtttttgta gttctttagt 7861 ttgtatgtct gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc 7921 cccttttctt ttaaaggcga taccgtcgag atccgttcac taatcgaatg gatctgtctc 7981 tgtctctctc tccaccttct tcttctattc cttcgggcct gtcgggtccc ctcggggttg 8041 ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc actatagaaa 8101 gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg aataatttta 8161 tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt 8221 ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta 8281 ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa 8341 tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga 8401 gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag 8461 ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt 8521 gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc 8581 tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga 8641 acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct 8701 ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat ttatataatt 8761 cacttctcca attgtccctc atatctcctc ctccaggtct gaagatcagc ggccggccgc 8821 ttgctgtgcg gtggtcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt 8881 ggtgtctttt atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa 8941 tgatctaagt tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct 9001 acagccttct gatgtttcta acaggccagg attaactgcg aatcgttcta gctccctgct 9061 tgcccatact atatgtttta atttatattt tttctttccc cctggcctta accgaatttt 9121 ttcccatcgc gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc 9181 cttctagcct ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct 9241 tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt 9301 cgctttcaag tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt 9361 ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact 9421 caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag 9481 gctcagatct ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca 9541 ggatctgagg gctcgccact ccccagtccc gcccaggcca cgcctccctg gaaagtcccc 9601 agcggaaagt cccttgtagc aagctcgata tcagcagttc ttgaagtact ccggatgcag 9661 ctctcgggcc acgtgatgaa atgctaggcg gctgtcaaac ctccactcta acacttctct 9721 ctccgggtca tccatcccat gcaggctcac agggtgtaac aagctggtgt tctctccttt 9781 attggcctct tctaccttat ctggctcaac tggtactagc ttgtagcacc atccaaaggt 9841 cagtggatat ctgacccctg gccctggtgt gtagttctgc taatcaggga agtagccttg 9901 tgtgtggtag atccacagat caaggatatc ttgtcttctt tgggagtgaa ttagcccttc 9961 ca //