LOCUS pGLuc_Mini-TK 4990 bp DNA circular 07-NOV-2008 DEFINITION Cloning vector pGLuc_Mini-TK, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pGLuc_Mini-TK ORGANISM Cloning vector pGLuc_Mini-TK other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 4990) AUTHORS Shea,C. and Tzertzinis,G. TITLE Direct Submission JOURNAL Submitted (07-NOV-2008) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..4990 /organism="Cloning vector pGLuc_Mini-TK" /mol_type="other DNA" misc_feature 12..68 /note="MCS (BglII-BamHI)" promoter 69..131 /note="minimal thymidine kinase promoter (PminiTK)" misc_RNA 140..149 /note="Kozak consensus sequence for GLuc" gene 146..703 /gene="GLuc" CDS 146..703 /gene="GLuc" /codon_start=1 /product="Gaussia princeps luciferase" /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADR GKLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKE SAQGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKW LPQRCATFASKIQGQVDKIKGAGGD" sig_peptide 146..196 /gene="GLuc" /note="secretory signal sequence" polyA_signal 722..770 /note="synthetic polyA" promoter 1308..1643 /note="SV40 promoter (clockwise)" gene 1695..2489 /gene="aph(3')-II" CDS 1695..2489 /gene="aph(3')-II" /note="neoR, kanR, nptII (confers resistance to neomycin and kanamycin)" /codon_start=1 /product="aminoglycoside phosphotransferase from Tn5" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(2882..2911) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)" rep_origin complement(3235..3823) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point) (old sequence errors)" gene complement(3994..4854) /gene="bla" CDS complement(3994..4854) /gene="bla" /note="ampR (confers resistance to ampicillin) (mutant V82I,A182V)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1145 a 1327 c 1306 g 1212 t ORIGIN 1 gacggatcgg gagatcttgg aattctgcag atatcctcga gcccaagctt ggtaccgagc 61 tcggatcctt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc 121 gacccgctta aaagatccag ccaccatggg agtcaaagtt ctgtttgccc tgatctgcat 181 cgctgtggcc gaggccaagc ccaccgagaa caacgaagac ttcaacatcg tggccgtggc 241 cagcaacttc gcgaccacgg atctcgatgc tgaccgcggg aagttgcccg gcaagaagct 301 gccgctggag gtgctcaaag agatggaagc caatgcccgg aaagctggct gcaccagggg 361 ctgtctgatc tgcctgtccc acatcaagtg cacgcccaag atgaagaagt tcatcccagg 421 acgctgccac acctacgaag gcgacaaaga gtccgcacag ggcggcatag gcgaggcgat 481 cgtcgacatt cctgagattc ctgggttcaa ggacttggag cccatggagc agttcatcgc 541 acaggtcgat ctgtgtgtgg actgcacaac tggctgcctc aaagggcttg ccaacgtgca 601 gtgttctgac ctgctcaaga agtggctgcc gcaacgctgt gcgacctttg ccagcaagat 661 ccagggccag gtggacaaga tcaagggggc cggtggtgac taagcggccg ctcgagcatg 721 catctagaaa taattcttac tgtcatgcca agtaagatgc ttttctgtgc tgcaatagca 781 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct 841 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 901 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 961 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt 1021 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 1081 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 1141 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 1201 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga 1261 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa 1321 gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 1381 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 1441 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 1501 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 1561 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 1621 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 1681 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 1741 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 1801 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 1861 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 1921 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 1981 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 2041 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 2101 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 2161 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 2221 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 2281 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 2341 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 2401 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 2461 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 2521 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 2581 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 2641 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 2701 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 2761 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 2821 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 2881 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 2941 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 3001 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 3061 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 3121 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 3181 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 3241 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 3301 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 3361 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 3421 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 3481 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 3541 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 3601 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 3661 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 3721 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 3781 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 3841 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 3901 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 3961 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 4021 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 4081 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 4141 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 4201 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 4261 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 4321 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 4381 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 4441 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 4501 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 4561 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 4621 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 4681 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 4741 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 4801 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 4861 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 4921 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 4981 acctgacgtc //