LOCUS pGLuc_Mini-TK_2 5028 bp DNA circular 24-SEP-2012 DEFINITION Cloning vector pGLuc_Mini-TK_2, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pGLuc_Mini-TK_2 ORGANISM Cloning vector pGLuc_Mini-TK_2 Unclassified. REFERENCE 1 (bases 1 to 5028) AUTHORS Tzertzinis,G. TITLE Direct Submission JOURNAL Submitted (24-SEP-2012) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..5028 /organism="Cloning vector pGLuc_Mini-TK_2" /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 712..760 /note="synthetic polyA" promoter 1346..1681 /note="SV40 promoter (clockwise)" gene 1733..2527 /gene="aph(3')-II" CDS 1733..2527 /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" rep_origin complement(3273..3861) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(4032..4892) /gene="bla" CDS complement(4032..4892) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1153 a 1328 c 1311 g 1236 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 caataaaata 721 tctttatttt cattacatct gtgtgttggt tttttgtgtg tctagaaata attcttactg 781 tcatgccaag taagatgctt ttctgtgctg caatagcagg catgctgggg atgcggtggg 841 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 901 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 961 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 1021 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt 1081 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 1141 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 1201 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 1261 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 1321 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg 1381 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 1441 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 1501 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 1561 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 1621 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 1681 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 1741 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 1801 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 1861 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 1921 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 1981 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 2041 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 2101 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 2161 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 2221 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 2281 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 2341 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 2401 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 2461 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 2521 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 2581 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 2641 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 2701 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 2761 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 2821 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 2881 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 2941 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 3001 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 3061 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 3121 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 3181 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 3241 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 3301 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 3361 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 3421 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt 3481 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 3541 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 3601 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 3661 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 3721 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 3781 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 3841 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 3901 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 3961 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 4021 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 4081 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 4141 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 4201 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 4261 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 4321 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 4381 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 4441 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 4501 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 4561 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 4621 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 4681 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 4741 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 4801 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 4861 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 4921 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 4981 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc //