LOCUS V028629 6667 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028629 VERSION V028629 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..2418 /label="USP14(NM_005151)" /note="USP14(NM_005151)" /gene="USP14" CDS 2431..2496 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2531..2755 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2801..3229 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3243..3572 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3584..4183 /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 4343..4464 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4513..4529) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4537..4553 /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(4561..4591) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4606..4627 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4915..5500) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5671..6531) /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(6532..6636) /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 gccaccatgc cgctctactc cgttactgta 961 aaatggggaa aggagaaatt tgaaggtgta gaattgaata cagatgaacc tccaatggta 1021 ttcaaggctc agctgtttgc gttgactgga gtccagcctg ccagacagaa agttatggtg 1081 aaaggaggaa cgctaaagga tgatgattgg ggaaacatca aaataaaaaa tggaatgact 1141 ctactaatga tggggtcagc agatgctctt ccagaagaac cctcagccaa aactgtcttc 1201 gtagaagaca tgacagaaga acagttagca tctgctatgg agttaccatg tggattgaca 1261 aaccttggta acacttgtta catgaatgcc acagttcagt gtattcgttc tgtgcctgaa 1321 ctcaaagatg cccttaaaag gtatgcaggt gccttgagag cttcagggga aatggcttca 1381 gcgcagtata ttactgcagc ccttagagat ttgtttgatt ccatggataa aacttcttcc 1441 agtattccac ctattattct actgcagttt ttgcacatgg ctttcccaca gtttgccgag 1501 aaaggtgaac aaggacagta tcttcaacag gatgctaatg aatgttggat acaaatgatg 1561 cgagtattgc aacagaaatt ggaagcaata gaggatgatt ctgttaaaga gacagactcc 1621 tcatctgcat cggcagcgac accttctaaa aagaaaagtt taatcgatca gttcttcggt 1681 gttgagtttg aaactaccat gaaatgtaca gaatctgaag aagaagaagt caccaaagga 1741 aaggaaaatc aacttcagct tagctgtttt atcaatcagg aagtcaagta tctttttaca 1801 ggacttaaat tgcgacttca ggaagaaatc accaaacagt ctccaacgtt gcaaagaaat 1861 gccttgtata tcaaatcttc caagatcagc cggctgcctg cttacttgac cattcagatg 1921 gttcgatttt tttataaaga gaaggaatct gtgaatgcca aagttcttaa ggatgttaaa 1981 tttcctctta tgttggatat gtatgaactg tgtacaccag aacttcaaga gaaaatggtg 2041 tcttttcgat ccaaattcaa ggatctagaa gataaaaaag tgaatcagca gccaaataca 2101 agtgacaaaa agagtagtcc ccagaaagaa gttaagtatg aacccttttc ttttgctgat 2161 gatattggct ccaataattg tggatactat gacttacaag cagtactaac acaccaggga 2221 aggtctagtt cttcaggtca ttatgtatca tgggtgaaaa ggaaacaaga tgaatggatt 2281 aagtttgatg atgacaaagt cagcatcgta acaccagaag atatcttacg gctttctggt 2341 ggtggagact ggcatatcgc ttacgttcta ctctatgggc ctcgcagagt tgaaataatg 2401 gaagaggaaa gtgaacagct cgagaccggt gactacaaag accatgacgg tgattataaa 2461 gatcatgaca tcgattacaa ggatgacgat gacaagtgac cggtaagttt aaaccgctga 2521 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 2581 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 2641 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 2701 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 2761 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 2821 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 2881 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 2941 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 3001 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 3061 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 3121 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggccatttcg 3181 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 3241 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3301 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 3361 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3421 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3481 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 3541 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggatgaccg agtacaagcc 3601 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc 3661 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt 3721 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 3781 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 3841 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 3901 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 3961 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 4021 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 4081 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga 4141 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgattcgaaa tgaccgacca 4201 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 4261 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 4321 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 4381 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 4441 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 4501 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 4561 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 4621 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 4681 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 4741 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 4801 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 4861 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 4921 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 4981 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 5041 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 5101 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 5161 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 5221 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 5281 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 5341 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 5401 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt 5461 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 5521 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 5581 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 5641 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 5701 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 5761 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 5821 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 5881 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 5941 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 6001 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 6061 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 6121 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 6181 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 6241 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 6301 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 6361 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 6421 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 6481 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 6541 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 6601 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 6661 tgacgtc //