LOCUS V024461 6584 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024461 VERSION V024461 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..2074 /label="TIMD4(NM_138379)" /note="TIMD4(NM_138379)" /gene="TIMD4" CDS 2081..2146 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2184..2408 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2454..2882 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2896..3225 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3292..4086 /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 4260..4381 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4430..4446) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4454..4470 /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(4478..4508) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4523..4544 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4832..5417) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5588..6448) /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(6449..6553) /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 atgtccaaag aacctctcat 961 tctctggctg atgattgagt tttggtggct ttacctgaca ccagtcactt cagagactgt 1021 tgtgacggag gttttgggtc accgggtgac tttgccctgt ctgtactcat cctggtctca 1081 caacagcaac agcatgtgct gggggaaaga ccagtgcccc tactccggtt gcaaggaggc 1141 gctcatccgc actgatggaa tgagggtgac ctcaagaaag tcagcaaaat atagacttca 1201 ggggactatc ccgagaggtg atgtctcctt gaccatctta aaccccagtg aaagtgacag 1261 cggtgtgtac tgctgccgca tagaagtgcc tggctggttc aacgatgtaa agataaacgt 1321 gcgcctgaat ctacagagag cctcaacaac cacgcacaga acagcaacca ccaccacacg 1381 cagaacaaca acaacaagcc ccaccaccac ccgacaaatg acaacaaccc cagctgcact 1441 tccaacaaca gtcgtgacca cacccgatct cacaaccgga acaccactcc agatgacaac 1501 cattgccgtc ttcacaacag caaacacgtg cctttcacta accccaagca cccttccgga 1561 ggaagccaca ggtcttctga ctcccgagcc ttctaaggaa gggcccatcc tcactgcaga 1621 atcagaaact gtcctcccca gtgattcctg gagtagtgtt gagtctactt ctgctgacac 1681 tgtcctgctg acatccaaag agtccaaagt ttgggatctc ccatcaacat cccacgtgtc 1741 aatgtggaaa acgagtgatt ctgtgtcttc tcctcagcct ggagcatctg atacagcagt 1801 tcctgagcag aacaaaacaa caaaaacagg acagatggat ggaataccca tgtcaatgaa 1861 gaatgaaatg cccatctccc aactactgat gatcatcgcc ccctccttgg gatttgtgct 1921 cttcgcattg tttgtggcgt ttctcctgag agggaaactc atggaaacct attgttcgca 1981 gaaacacaca aggctagact acattggaga tagtaaaaat gtcctcaatg acgtgcagca 2041 tggaagggaa gacgaagacg gcctttttac cctcctcgag gactacaaag accatgacgg 2101 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 2161 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 2221 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 2281 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 2341 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 2401 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 2461 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 2521 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 2581 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2641 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2701 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2761 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 2821 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 2881 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 2941 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 3001 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 3061 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 3121 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 3181 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 3241 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 3301 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 3361 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 3421 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 3481 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 3541 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 3601 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3661 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3721 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3781 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 3841 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3901 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3961 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 4021 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 4081 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 4141 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 4201 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 4261 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4321 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4381 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 4441 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 4501 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4561 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4621 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4681 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4741 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4801 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4861 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 5041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5161 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5221 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 5281 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5341 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 5401 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 5461 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 5521 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 5581 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 5641 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5701 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 5761 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 5821 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 5881 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 5941 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 6001 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 6061 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6121 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 6181 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 6241 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 6301 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 6361 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 6421 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 6481 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 6541 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //