LOCUS V042544 6841 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042544 VERSION V042544 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 925..2592 /label="PDPK1(NM_002613)" /note="PDPK1(NM_002613)" /gene="PDPK1" CDS 2605..2670 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2705..2929 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2975..3403 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3417..3746 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3758..4357 /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 4517..4638 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4687..4703) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4711..4727 /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(4735..4765) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4780..4801 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5089..5674) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5845..6705) /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(6706..6810) /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 gagaattcgc caccatggcc aggaccacca gccagctgta tgacgccgtg 961 cccatccagt ccagcgtggt gttatgttcc tgcccatccc catcaatggt gaggacccag 1021 actgagtcca gcacgccccc tggcattcct ggtggcagca ggcagggccc cgccatggac 1081 ggcactgcag ccgagcctcg gcccggcgcc ggctccctgc agcatgccca gcctccgccg 1141 cagcctcgga agaagcggcc tgaggacttc aagtttggga aaatccttgg ggaaggctct 1201 ttttccacgg ttgtcctggc tcgagaactg gcaacctcca gagaatatgc gattaaaatt 1261 ctggagaagc gacatatcat aaaagagaac aaggtcccct atgtaaccag agagcgggat 1321 gtcatgtcgc gcctggatca ccccttcttt gttaagcttt acttcacatt tcaggacgac 1381 gagaagctgt atttcggcct tagttatgcc aaaaatggag aactacttaa atatattcgc 1441 aaaatcggtt cattcgatga gacctgtacc cgattttaca cggctgagat tgtgtctgct 1501 ttagagtact tgcacggcaa gggcatcatt cacagggacc ttaaaccgga aaacattttg 1561 ttaaatgaag atatgcacat ccagatcaca gattttggaa cagcaaaagt cttatcccca 1621 gagagcaaac aagccagggc caactcattc gtgggaacag cgcagtacgt ttctccagag 1681 ctgctcacgg agaagtccgc ctgtaagagt tcagaccttt gggctcttgg atgcataata 1741 taccagcttg tggcaggact cccaccattc cgagctggaa acgagtatct tatatttcag 1801 aagatcatta agttggaata tgactttcca gaaaaattct tccctaaggc aagagacctc 1861 gtggagaaac ttttggtttt agatgccaca aagcggttag gctgtgagga aatggaagga 1921 tacggacctc ttaaagcaca cccgttcttc gagtccgtca cgtgggagaa cctgcaccag 1981 cagacgcctc cgaagctcac cgcttacctg ccggctatgt cggaagacga cgaggactgc 2041 tatggcaatt atgacaatct cctgagccag tttggctgca tgcaggtgtc ttcgtcctcc 2101 tcctcacact ccctgtcagc ctccgacacg ggcctgcccc agaggtcagg cagcaacata 2161 gagcagtaca ttcacgatct ggactcgaac tcctttgaac tggacttaca gttttccgaa 2221 gatgagaaga ggttgttgtt ggagaagcag gctggcggaa acccttggca ccagtttgta 2281 gaaaataatt taatactaaa gatgggccca gtggataagc ggaagggttt atttgcaaga 2341 cgacgacagc tgttgctcac agaaggacca catttatatt atgtggatcc tgtcaacaaa 2401 gttctgaaag gtgaaattcc ttggtcacaa gaacttcgac cagaggccaa gaattttaaa 2461 actttctttg tccacacgcc taacaggacg tattatctga tggaccccag cgggaacgca 2521 cacaagtggt gcaggaagat ccaggaggtt tggaggcagc gataccagag ccacccggac 2581 gccgctgtgc agctcgagac cggtgactac aaagaccatg acggtgatta taaagatcat 2641 gacatcgatt acaaggatga cgatgacaag tgaccggtaa gtttaaaccg ctgatcagcc 2701 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 2761 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 2821 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 2881 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 2941 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 3001 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 3061 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3121 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 3181 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 3241 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 3301 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggccat ttcggcctat 3361 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 3421 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3481 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 3541 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 3601 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 3661 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 3721 tttttggagg cctaggcttt tgcaaaaagc tcccgggatg accgagtaca agcccacggt 3781 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc 3841 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga 3901 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga 3961 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt 4021 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca 4081 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt 4141 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt 4201 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct 4261 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc 4321 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgattc gaaatgaccg accaagcgac 4381 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 4441 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 4501 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4561 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4621 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 4681 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 4741 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 4801 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 4861 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 4921 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4981 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 5041 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 5101 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 5161 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 5221 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 5281 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 5341 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 5401 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 5461 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 5521 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 5581 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa 5641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6841 c //