LOCUS V018437 7589 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018437 VERSION V018437 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 671..2269 /label="PHGDH(NM_006623)" /note="PHGDH(NM_006623)" /gene="PHGDH" CDS 2276..2341 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2365..2953 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3019..3067 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3269..3697 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3711..4040 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4107..4901 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 5077..5198 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5247..5263) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5271..5287 /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(5295..5325) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5340..5361 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5649..6237) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6408..7268) /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(7269..7373) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgccacc atggcttttg caaatctgcg gaaagtgctc atcagtgaca gcctggaccc 721 ttgctgccgg aagatcttgc aagatggagg gctgcaggtg gtggaaaagc agaaccttag 781 caaagaggag ctgatagcgg agctgcagga ctgtgaaggc cttattgttc gctctgccac 841 caaggtgacc gctgatgtca tcaacgcagc tgagaaactc caggtggtgg gcagggctgg 901 cacaggtgtg gacaatgtgg atctggaggc cgcaacaagg aagggcatct tggttatgaa 961 cacccccaat gggaacagcc tcagtgccgc agaactcact tgtggaatga tcatgtgcct 1021 ggccaggcag attccccagg cgacggcttc gatgaaggac ggcaaatggg agcggaagaa 1081 gttcatggga acagagctga atggaaagac cctgggaatt cttggcctgg gcaggattgg 1141 gagagaggta gctacccgga tgcagtcctt tgggatgaag actatagggt atgaccccat 1201 catttcccca gaggtctcgg cctcctttgg tgttcagcag ctgcccctgg aggagatctg 1261 gcctctctgt gatttcatca ctgtgcacac tcctctcctg ccctccacga caggcttgct 1321 gaatgacaac acctttgccc agtgcaagaa gggggtgcgt gtggtgaact gtgcccgtgg 1381 agggatcgtg gacgaaggcg ccctgctccg ggccctgcag tctggccagt gtgccggggc 1441 tgcactggac gtgtttacgg aagagccgcc acgggaccgg gccttggtgg accatgagaa 1501 tgtcatcagc tgtccccacc tgggtgccag caccaaggag gctcagagcc gctgtgggga 1561 ggaaattgct gttcagttcg tggacatggt gaaggggaaa tctctcacgg gggttgtgaa 1621 tgcccaggcc cttaccagtg ccttctctcc acacaccaag ccttggattg gtctggcaga 1681 agctctgggg acactgatgc gagcctgggc tgggtccccc aaagggacca tccaggtgat 1741 aacacaggga acatccctga agaatgctgg gaactgccta agccccgcag tcattgtcgg 1801 cctcctgaaa gaggcttcca agcaggcgga tgtgaacttg gtgaacgcta agctgctggt 1861 gaaagaggct ggcctcaatg tcaccacctc ccacagccct gctgcaccag gggagcaagg 1921 cttcggggaa tgcctcctgg ccgtggccct ggcaggcgcc ccttaccagg ctgtgggctt 1981 ggtccaaggc actacgcctg tactgcaggg gctcaatgga gctgtcttca ggccagaagt 2041 gcctctccgc agggacctgc ccctgctcct attccggact cagacctctg accctgcaat 2101 gctgcctacc atgattggcc tcctggcaga ggcaggcgtg cggctgctgt cctaccagac 2161 ttcactggtg tcagatgggg agacctggca cgtcatgggc atctcctcct tgctgcccag 2221 cctggaagcg tggaagcagc atgtgactga agccttccag ttccacttcc tcgaggacta 2281 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa 2341 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg 2401 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 2461 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 2521 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 2581 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 2641 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 2701 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag 2761 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 2821 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 2881 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 2941 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg 3001 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg 3061 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga 3121 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg 3181 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg 3241 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 3301 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 3361 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 3421 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 3481 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 3541 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 3601 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta 3661 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 3721 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3781 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3841 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3901 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3961 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 4021 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 4081 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 4141 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 4201 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 4261 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 4321 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 4381 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 4441 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 4501 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 4561 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 4621 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 4681 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 4741 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 4801 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 4861 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc 4921 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 4981 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 5041 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 5101 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 5161 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 5221 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 5281 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 5341 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 5401 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 5461 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 5521 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 5581 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 5641 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 5701 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 5761 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 5821 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc 5881 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 5941 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 6001 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 6061 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 6121 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 6181 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 6241 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 6301 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 6361 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 6421 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 6481 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 6541 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 6601 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 6661 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 6721 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 6781 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 6841 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 6901 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 6961 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 7021 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 7081 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 7141 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 7201 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 7261 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 7321 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 7381 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga 7441 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg 7501 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga 7561 ccgacaattg catgaagaat ctgcttagg //