LOCUS V032055 6104 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032055 VERSION V032055 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..1594 /label="Fgf19(NM_130753)" /note="Fgf19(NM_130753)" /gene="Fgf19" CDS 1601..1666 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1704..1928 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1974..2402 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2416..2745 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2812..3606 /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 3780..3901 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3950..3966) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3974..3990 /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(3998..4028) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4043..4064 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4352..4937) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5108..5968) /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(5969..6073) /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 atggcgagaa agtggagtgg 961 gcgtattgtg gcccgagctc tggtcctggc cactctgtgg ctggccgtgt ctgggcgtcc 1021 cctggtccag caatcccagt ctgtgtcgga tgaaggtcca ctctttctct atggctgggg 1081 caagattacc cgcctgcagt acctgtactc tgctggtccc tacgtctcca actgcttcct 1141 gcgtatccgg agtgacggct ctgtggactg cgaggaggac cagaacgaac gaaatctgtt 1201 ggagttccgc gcggttgctc tgaagacaat tgccatcaag gacgtcagca gcgtgcggta 1261 cctctgcatg agcgccgacg gcaagatata cgggctgatt cgctactcgg aggaagactg 1321 taccttcagg gaggaaatgg actgtttggg ctacaaccag tacaggtcca tgaagcacca 1381 cctccacatc atcttcatca aggccaagcc cagagagcag ctccagggcc agaaaccttc 1441 aaactttatc cccatatttc accggtcttt ctttgaatcc acggaccagc tgaggtctaa 1501 aatgttctct ctgcccctgg agagcgacag catggatccg ttcagaatgg tggaggatgt 1561 ggaccaccta gtgaagagtc ccagcttcca gaaactcgag gactacaaag accatgacgg 1621 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 1681 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 1741 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 1801 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 1861 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 1921 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 1981 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 2041 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 2101 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2161 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2221 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2281 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 2341 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 2401 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 2461 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 2521 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 2581 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 2641 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 2701 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 2761 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 2821 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 2881 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 2941 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 3001 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 3061 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 3121 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3181 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3241 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3301 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 3361 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3421 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3481 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 3541 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 3601 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 3661 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 3721 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 3781 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 3841 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 3901 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 3961 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 4021 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4081 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4141 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4201 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4261 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4321 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4381 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4441 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4501 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 4561 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 4621 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 4681 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 4741 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 4801 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 4861 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 4921 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 4981 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 5041 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 5101 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 5161 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5221 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 5281 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 5341 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 5401 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 5461 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 5521 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 5581 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 5641 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 5701 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 5761 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 5821 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 5881 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 5941 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 6001 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 6061 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //