LOCUS V025352 6074 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025352 VERSION V025352 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..1564 /label="FGF6(NM_020996)" /note="FGF6(NM_020996)" /gene="FGF6" CDS 1571..1636 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1674..1898 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1944..2372 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2386..2715 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2782..3576 /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 3750..3871 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3920..3936) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3944..3960 /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(3968..3998) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4013..4034 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4322..4907) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5078..5938) /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(5939..6043) /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 atggccctgg gacagaaact 961 gttcatcact atgtcccggg gagcaggacg tctgcagggc acgctgtggg ctctcgtctt 1021 cctaggcatc ctagtgggca tggtggtgcc ctcgcctgca ggcacccgtg ccaacaacac 1081 gctgctggac tcgaggggct ggggcaccct gctgtccagg tctcgcgccg ggctagctgg 1141 agagattgcc ggggtgaact gggaaagtgg ctatttggtg gggatcaagc ggcagcggag 1201 gctctactgc aacgtgggca tcggctttca cctccaggtg ctccccgacg gccggatcag 1261 cgggacccac gaggagaacc cctacagcct gctggaaatt tccactgtgg agcgaggcgt 1321 ggtgagtctc tttggagtga gaagtgccct cttcgttgcc atgaacagta aaggaagatt 1381 gtacgcaacg cccagcttcc aagaagaatg caagttcaga gaaaccctcc tgcccaacaa 1441 ttacaatgcc tacgagtcag acttgtacca agggacctac attgccctga gcaaatacgg 1501 acgggtaaag cggggcagca aggtgtcccc gatcatgact gtcactcatt tccttcccag 1561 gatcctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 1621 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 1681 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 1741 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 1801 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 1861 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 1921 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 1981 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 2041 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 2101 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 2161 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 2221 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 2281 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 2341 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 2401 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2461 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2521 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 2581 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 2641 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 2701 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 2761 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 2821 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 2881 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 2941 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 3001 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 3061 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 3121 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 3181 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 3241 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 3301 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 3361 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 3421 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 3481 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 3541 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 3601 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 3661 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 3721 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 3781 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 3841 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 3901 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 3961 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 4021 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 4081 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 4141 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 4201 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4261 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4321 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4381 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 4441 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 4501 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 4561 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 4621 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 4681 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 4741 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 4801 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt 4861 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 4921 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 4981 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 5041 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 5101 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 5161 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 5221 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 5281 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 5341 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 5401 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 5461 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 5521 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 5581 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 5641 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 5701 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5761 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5821 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5881 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 5941 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 6001 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 6061 tgccacctga cgtc //