LOCUS V018355 7475 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018355 VERSION V018355 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 941..2932 /label="CHFR(NM_001161344)" /note="CHFR(NM_001161344)" /gene="CHFR" CDS 2972..3037 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3075..3299 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3345..3773 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3787..4116 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4183..4977 /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 5151..5272 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5321..5337) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5345..5361 /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(5369..5399) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5414..5435 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5723..6308) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6479..7339) /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(7340..7444) /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 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggagcggc ccgaggaagg 961 caagcagtcg ccgccgccgc agccctgggg acggctcctg cgtctgggcg cggaggaggg 1021 cgagccgcac gtcctcctga ggaagcggga gtggaccatc gggcggagac gaggttgcga 1081 cctttccttc cccagcaata aactggtctc tggagatcac tgtagaattg tagtggatga 1141 aaaatcaggt caggtgacac tggaagatac cagcaccagt ggaacagtga ttaacaagct 1201 gaaggttgtt aagaagcaga catgcccttt acagactggg gatgtcatct acttggtgta 1261 caggaagaat gaaccggaac acaacgtggc atacctctat gaatctttaa gtgaaaagca 1321 aggcatgaca caagaatcct ttgaagctaa caaggaaaat gtgttccatg ggaccaaaga 1381 tacctcaggt gcaggtgcag ggcgaggggc cgatccccgg gtccctccgt cgtcgcccgc 1441 cactcaggtg tgctttgagg aaccacagcc atcaacatcg acgtcagacc tcttccccac 1501 agcctcggcc tcttccacgg agccttctcc tgcagggcga gagcgttcct ccagttgtgg 1561 gtctgggggt ggtggcatct cccctaaagg aagtggtccc tctgtggcaa gtgatgaagt 1621 ctccagcttt gcctcagctc tcccagacag aaagactgcg tccttttcgt cgttggaacc 1681 ccaggatcag gaggatttgg agcccgtgaa gaagaaaatg agaggagatg gggaccttga 1741 cctgaacggg cagttgttgg tcgcacaacc gcgtagaaat gcccaaaccg tccacgagga 1801 cgtcagagca gcggctggga agccagacaa gatggaggag acgctgacat gcatcatctg 1861 ccaggacctg ctgcacgact gcgtgagttt gcagccctgc atgcacacgt tctgcgcggc 1921 ttgctactcg ggctggatgg agcgctcgtc cctgtgtcct acctgccgct gtcccgtgga 1981 gcggatctgt aaaaaccaca tcctcaacaa cctcgtggaa gcatacctca tccagcatcc 2041 agacaagagt cgcagtgaag aagatgtgca aagtatggat gccaggaata aaatcactca 2101 agacatgctg cagcccaaag tcaggcggtc tttttctgat gaagaaggga gttcagagga 2161 cctgctggag ctgtcagacg ttgacagtga gtcctcagac attagccagc catacgtcgt 2221 gtgccggcag tgtcctgagt acagaaggca ggcggcgcag cctccccact gcccagcacc 2281 cgagggcgag ccaggagccc cacaggccct gggggatgca ccctccacgt ccgtcagcct 2341 gacgacagca gtccaggatt acgtgtgccc tctgcaagga agccacgccc tgtgcacctg 2401 ctgcttccag cccatgcccg accggagagc ggagcgcgag caggacccgc gtgtcgcccc 2461 tcagcagtgt gcggtctgcc tgcagccttt ctgccacctg tactggggct gcacccggac 2521 cggctgctac ggctgcctgg ccccgttttg tgagctcaac ctgggtgaca agtgtctgga 2581 cggcgtgctg aacaacaaca gctacgagtc agacatcctg aagaattacc tggcaaccag 2641 aggtttgaca tggaaaaaca tgttgaccga gagcctcgtg gctctccagc ggggagtgtt 2701 tctgctgtct gattacagag tcacgggaga caccgttctg tgttactgct gtggcctgcg 2761 cagcttccgt gagctgacct atcagtatcg gcagaacatt cctgcttccg agttgccagt 2821 ggccgtaaca tcccgtcctg actgctactg gggccgtaac tgccgcactc aggtgaaagc 2881 tcaccacgcc atgaaattca atcatatctg tgaacagaca aggttcaaaa acgaattctg 2941 cagatatcca gcacagtggc ggccgctcga ggactacaaa gaccatgacg gtgattataa 3001 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg 3061 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 3121 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 3181 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 3241 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 3301 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 3361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 3421 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 3481 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 3541 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 3601 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 3661 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 3721 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 3781 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 3841 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 3961 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 4021 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 4081 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 4141 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 4201 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 4261 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 4321 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 4381 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 4441 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 4501 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 4561 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 4621 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 4681 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 4741 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 4801 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 4861 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 4921 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 4981 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 5041 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 5101 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 5161 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 5221 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 5281 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 5341 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 5401 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 5461 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 5521 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 5581 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 5641 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 5701 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 5761 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 5821 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 5881 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 5941 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 6001 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 6061 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 6121 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 6181 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 6241 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 6301 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 6361 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 6421 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 6481 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 6541 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 6601 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 6661 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 6721 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 6781 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 6841 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 6901 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 6961 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 7021 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 7081 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 7141 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 7201 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 7261 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 7321 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 7381 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 7441 cgcgcacatt tccccgaaaa gtgccacctg acgtc //