LOCUS V020671 6047 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020671 VERSION V020671 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 905..1798 /label="GOLPH3(NM_022130)" /note="GOLPH3(NM_022130)" /gene="GOLPH3" CDS 1811..1876 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1911..2135 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2181..2609 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2623..2952 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2964..3563 /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 3723..3844 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3893..3909) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3917..3933 /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(3941..3971) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3986..4007 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4295..4880) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5051..5911) /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(5912..6016) /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 caccatgacc tcgctgaccc agcgcagctc cggcctggtg cagcggcgca ccgaggcctc 961 ccgcaacgcc gccgacaagg agcgggcggc gggcggcggc gccggcagca gcgaggacga 1021 cgcgcagagc cgccgcgacg agcaggacga cgacgacaag ggcgactcca aggaaacgcg 1081 gctgaccctg atggaggaag tgctcctgct gggcctcaag gaccgcgagg gttacacatc 1141 attttggaat gactgtatat catctggatt acgtggctgt atgttaattg aattagcatt 1201 gagaggaagg ttacaactag aggcttgtgg aatgagacgt aaaagtctat taacaagaaa 1261 ggtaatctgt aagtcagatg ctccaacagg ggatgttctt cttgatgaag ctctgaagca 1321 tgttaaggaa actcagcctc cagaaacggt ccagaactgg attgaattac ttagtggtga 1381 gacatggaat ccattaaaat tgcattatca gttaagaaat gtacgggaac gattagctaa 1441 aaacctggtg gaaaagggtg tattgacaac agagaaacag aacttcctac tttttgacat 1501 gacaacacat cccctcacca ataacaacat taagcagcgc ctcatcaaga aagtacagga 1561 agccgttctt gacaaatggg tgaatgaccc tcaccgcatg gacaggcgct tgctggccct 1621 catttacctg gctcatgcct cggacgtcct ggagaatgct tttgctcctc ttctggacga 1681 gcagtatgat ttggctacca agagagtgcg gcagcttctc gacttagacc ctgaagtgga 1741 atgtctgaag gccaacacca atgaggttct gtgggcggtg gtggcggcgt tcaccaagct 1801 cgagaccggt gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa 1861 ggatgacgat gacaagtgac cggtaagttt aaaccgctga tcagcctcga ctgtgccttc 1921 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 1981 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 2041 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 2101 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg 2161 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 2221 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 2281 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 2341 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 2401 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 2461 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 2521 ggtctattct tttgatttat aagggatttt ggccatttcg gcctattggt taaaaaatga 2581 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 2641 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 2701 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 2761 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 2821 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 2881 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 2941 ggcttttgca aaaagctccc gggatgaccg agtacaagcc cacggtgcgc ctcgccaccc 3001 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca 3061 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct 3121 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg 3181 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc 3241 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc 3301 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg 3361 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc 3421 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc 3481 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca 3541 tgacccgcaa gcccggtgcc tgattcgaaa tgaccgacca agcgacgccc aacctgccat 3601 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 3661 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 3721 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 3781 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 3841 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 3901 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 3961 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4021 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 4081 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 4141 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4201 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4261 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4321 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4381 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4441 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 4501 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 4561 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 4621 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 4681 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 4741 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 4801 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc 4861 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 4921 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 4981 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 5041 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 5101 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 5161 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 5221 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 5281 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 5341 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 5401 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 5461 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 5521 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 5581 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 5641 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 5701 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 5761 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 5821 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 5881 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 5941 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6001 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //