LOCUS V038707 6832 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038707 VERSION V038707 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 937..1653 /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 1690..2583 /label="Stx4a(NM_009294)" /note="Stx4a(NM_009294)" /gene="Stx4a" CDS 2596..2661 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2696..2920 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2966..3394 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3408..3737 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3749..4348 /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" polyA_signal 4508..4629 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4678..4694) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4702..4718 /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(4726..4756) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4771..4792 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5080..5665) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5836..6696) /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(6697..6801) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 aagctttcta gagaattcga tatcggatcc gccaccatgg tgagcaaggg cgaggagctg 961 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 1021 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 1081 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc 1141 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 1201 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 1261 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 1321 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 1381 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc 1441 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 1501 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg 1561 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 1621 gggatcactc tcggcatgga cgagctgtac aagggcggtg gaggcagtgg tggaggcggt 1681 agcggtacca tgcgcgacag gacccacgag ttgaggcagg gggataacat ctcagacgat 1741 gaagatgagg ttcgagtcgc gctggtggtg cactcaggtg ctgcccggtt gggcagcccg 1801 gacgacgagt tcttccagaa ggtgcagaca attcggcaga ctatggccaa actggagagt 1861 aaagtccggg agttggagaa acagcaggtc accatcctgg ccacgcctct tcccgaggag 1921 agcatgaagc agggcctgca gaacctgcga gaggagatca aacagctggg gagagaggtc 1981 cgggcgcagc taaaagccat agagccccag aaggaggaag ctgatgagaa ttacaactca 2041 gtcaacacaa ggatgaagaa aacccagcat ggagtcctgt cccagcaatt tgtcgagctc 2101 atcaacaagt gcaactcaat gcagtccgaa taccgagaga agaatgtgga gcggatccgg 2161 aggcagctga agatcaccaa tgctggaatg gtgtctgacg aggagctgga acagatgctg 2221 gacagtgggc agagtgaggt gtttgtgtct aatatcctga aggacacgca ggtgactcgg 2281 caggccctga atgagatctc tgcgcggcac agtgagatcc agcagttgga gcgcagtatc 2341 cgagagctcc atgagatctt tacgtttcta gctacggagg tggagatgca gggggagatg 2401 atcaaccgca tcgagaagaa catcctgagc tcggccgact acgtggaacg tgggcaagag 2461 cacgtcaaga tagccctaga gaatcagaag aaggcgagga agaaaaaagt catgattgcc 2521 atctgtgtct ctgtcactgt tctcatcttg gctgtcatca ttggcatcac cataaccgtt 2581 ggactcgaga ccggtgacta caaagaccat gacggtgatt ataaagatca tgacatcgat 2641 tacaaggatg acgatgacaa gtgaccggta agtttaaacc gctgatcagc ctcgactgtg 2701 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 2761 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 2821 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 2881 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 2941 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 3001 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3061 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3121 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 3181 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 3241 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 3301 atctcggtct attcttttga tttataaggg attttggcca tttcggccta ttggttaaaa 3361 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 3421 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3481 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3541 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 3601 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 3661 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 3721 gcctaggctt ttgcaaaaag ctcccgggat gaccgagtac aagcccacgg tgcgcctcgc 3781 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc 3841 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga 3901 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc 3961 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat 4021 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg 4081 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc 4141 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc 4201 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta 4261 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg 4321 gtgcatgacc cgcaagcccg gtgcctgatt cgaaatgacc gaccaagcga cgcccaacct 4381 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 4441 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 4501 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 4561 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 4621 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 4681 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 4741 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 4801 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 4861 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 4921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 4981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 5041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 5101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 5161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 5221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 5281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 5341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 5401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 5461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 5521 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat 5641 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 5701 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 5761 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 5821 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 5881 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 5941 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 6001 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 6061 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 6121 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 6181 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 6241 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 6301 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 6361 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 6421 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 6481 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 6541 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 6601 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 6661 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 6721 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 6781 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //