LOCUS V033676 5864 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033676 VERSION V033676 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..1354 /label="FAM136A(NM_032822)" /note="FAM136A(NM_032822)" /gene="FAM136A" CDS 1361..1426 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1464..1688 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1734..2162 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2176..2505 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2572..3366 /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 3540..3661 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3710..3726) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3734..3750 /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(3758..3788) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3803..3824 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4112..4697) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4868..5728) /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(5729..5833) /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 atggctgagc tgcagcagct 961 ccgggtgcag gaggcggtgg agtccatggt gaagagtctg gaaagagaga acatccggaa 1021 gatgcagggt ctcatgttcc ggtgcagcgc cagctgttgt gaggacagcc aggcctccat 1081 gaagcaggtg caccagtgca tcgagcgctg ccatgtgcct ctggctcaag cccaggcttt 1141 ggtcaccagt gagctggaga agttccagga ccgcctggcc cggtgcacca tgcattgcaa 1201 cgacaaagcc aaagattcaa tagatgctgg gagtaaggag cttcaggtga agcagcagct 1261 ggacagttgt gtgaccaagt gtgtggatga ccacatgcac ctcatcccaa ctatgaccaa 1321 gaagatgaag gaggctctct tatcaattgg aaaactcgag gactacaaag accatgacgg 1381 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 1441 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 1501 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 1561 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 1621 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 1681 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 1741 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 1801 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 1861 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 1921 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 1981 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2041 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 2101 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 2161 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 2221 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 2281 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 2341 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 2401 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 2461 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 2521 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 2581 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 2641 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 2701 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 2761 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 2821 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 2881 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 2941 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3001 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3061 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 3121 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3181 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3241 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 3301 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 3361 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 3421 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 3481 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 3541 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 3601 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 3661 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 3721 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 3781 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 3841 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 3901 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 3961 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4021 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4141 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 4321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 4381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 4441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 4501 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 4561 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 4621 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 4681 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 4741 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 4801 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 4861 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 4921 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 4981 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 5041 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 5101 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 5161 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 5221 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 5281 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 5341 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 5401 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 5461 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 5521 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 5581 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 5641 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 5701 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 5761 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 5821 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //