LOCUS pCLuc_Mini-TK_2 6131 bp DNA circular 09-JUL-2010 DEFINITION Cloning vector pCLuc_Mini-TK_2, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pCLuc_Mini-TK_2 ORGANISM Cloning vector pCLuc_Mini-TK_2 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6131) AUTHORS Shea,C. and Tzertzinis,G. TITLE Direct Submission JOURNAL Submitted (09-JUL-2010) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6131 /organism="Cloning vector pCLuc_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 139..148 /note="Kozak consensus sequence for CLuc" gene 145..1806 /gene="CLuc" CDS 145..1806 /gene="CLuc" /codon_start=1 /product="Cypridina noctiluca luciferase" /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECID SSCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTY VLGQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADP IIANPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLC GDLKMMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPI NFYYYTISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQ FQGPCKEILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVG GEAVSVPYSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGIC GNYNQDFSDDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHK PDRVERCMYEYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK" polyA_signal 1815..1863 /note="synthetic polyA" promoter 2449..2784 /note="SV40 promoter (clockwise)" gene 2836..3630 /gene="aph(3')-II" CDS 2836..3630 /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" polyA_signal 3804..3934 /note="SV40 early polyA" promoter complement(4023..4052) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)" rep_origin complement(4376..4964) /note="pUC19 origin of replication () (RNAII -35 to RNA/DNA switch point) (old sequence errors)" gene complement(5135..5995) /gene="bla" CDS complement(5135..5995) /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 1496 a 1569 c 1546 g 1520 t ORIGIN 1 gacggatcgg gagatcttgg aattctgcag atatcctcga gcccaagctt ggtaccgagc 61 tcggatcctt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc 121 gacccgctta aaagatccgc caccatgaag accttaattc ttgccgttgc attagtctac 181 tgcgccactg ttcattgcca ggactgtcct tacgaacctg atccaccaaa cacagttcca 241 acttcctgtg aagctaaaga aggagaatgt attgatagca gctgtggcac ctgcacgaga 301 gacatactat cagatggact gtgtgaaaat aaaccaggaa aaacatgttg ccgaatgtgt 361 cagtatgtaa ttgaatgcag agtagaggcc gcaggatggt ttagaacatt ctatggaaag 421 agattccagt tccaggaacc tggtacatac gtgttgggtc aaggaaccaa gggcggcgac 481 tggaaggtgt ccatcaccct ggagaacctg gatggaacca agggggctgt gctgaccaag 541 acaagactgg aagtggctgg agacatcatt gacatcgctc aagctactga gaatcccatc 601 actgtaaacg gtggagctga ccctatcatc gccaacccgt acaccatcgg cgaggtcacc 661 atcgctgttg ttgagatgcc aggcttcaac atcaccgtca ttgagttctt caaactgatc 721 gtgatcgaca tcctcggagg aagatctgta agaatcgccc cagacacagc aaacaaagga 781 atgatctctg gcctctgtgg agatcttaaa atgatggaag atacagactt cacttcagat 841 ccagaacaac tcgctattca gcctaagatc aaccaggagt ttgacggttg tccactctat 901 ggaaatcctg atgacgttgc atactgcaaa ggtcttctgg agccgtacaa ggacagctgc 961 cgcaacccca tcaacttcta ctactacacc atctcctgcg ccttcgcccg ctgtatgggt 1021 ggagacgagc gagcctcaca cgtgctgctt gactacaggg agacgtgcgc tgctcccgaa 1081 actagaggaa cctgcgtttt gtctggacat actttctacg atacatttga caaagcaaga 1141 taccaattcc agggtccctg caaggagatt cttatggccg ccgactgttt ctggaacact 1201 tgggatgtga aggtttcaca caggaatgtt gactcttaca ctgaagtaga gaaagtacga 1261 atcaggaaac aatcgactgt agtagaactc attgttgatg gaaaacagat tctggttgga 1321 ggagaagccg tgtccgtccc gtacagctct cagaacactt ccatctactg gcaagatggt 1381 gacatactga ctacagccat cctacctgaa gctctggtgg tcaagttcaa cttcaagcaa 1441 ctgctcgtcg tacatattag agatccattc gatggtaaga cttgcggtat ttgcggtaac 1501 tacaaccagg atttcagtga tgattctttt gatgctgaag gagcctgtga tctgaccccc 1561 aacccaccgg gatgcaccga agaacagaaa cctgaagctg aacgactctg caatagtctc 1621 ttcgccggtc aaagtgatct tgatcagaaa tgtaacgtgt gccacaagcc tgaccgtgtc 1681 gaacgatgca tgtacgagta ttgcctgagg ggacaacagg gtttctgtga ccacgcatgg 1741 gagttcaaga aagaatgcta cataaagcat ggagacaccc tagaagtacc agatgaatgc 1801 aaataggcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt 1861 gtgtctagaa ataattctta ctgtcatgcc aagtaagatg cttttctgtg ctgcaatagc 1921 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 1981 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 2041 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 2101 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct 2161 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 2221 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 2281 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 2341 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg 2401 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 2461 agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 2521 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 2581 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 2641 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 2701 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 2761 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga 2821 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 2881 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 2941 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 3001 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 3061 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 3121 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 3181 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 3241 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 3301 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 3361 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 3421 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 3481 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 3541 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 3601 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 3661 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 3721 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 3781 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 3841 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 3901 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 3961 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 4021 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 4081 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 4141 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 4201 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 4261 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 4321 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 4381 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 4441 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 4501 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 4561 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 4621 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 4681 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 4741 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 4801 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 4861 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 4921 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 4981 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5041 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5101 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 5161 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 5221 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 5281 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 5341 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 5401 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 5461 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 5521 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 5581 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 5641 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 5701 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 5761 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 5821 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 5881 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 5941 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6001 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6061 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6121 cacctgacgt c //