LOCUS pTK-CLuc 7000 bp DNA circular 09-JUL-2010 DEFINITION Cloning vector pTK-CLuc, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pTK-CLuc ORGANISM Cloning vector pTK-CLuc other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 7000) 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..7000 /organism="Cloning vector pTK-CLuc" /mol_type="other DNA" promoter 18..770 /note="HSV TK promoter (clockwise)" misc_RNA 795..804 /note="Kozak consensus sequence for CLuc" gene 801..2462 /gene="CLuc" CDS 801..2462 /gene="CLuc" /codon_start=1 /product="Cypridina noctiluca luciferase" /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECID SSCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTY VLGQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADP IIANPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLC GDLKMMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPI NFYYYTISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQ FQGPCKEILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVG GEAVSVPYSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGIC GNYNQDFSDDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHK PDRVERCMYEYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK" sig_peptide 801..854 /gene="CLuc" /note="General Secretory Pathway signal peptide" misc_feature 2463..2494 /note="MCS (NotI-XbaI)" polyA_signal 2495..2725 /note="SV40 polyA" promoter 3318..3653 /note="SV40 promoter (clockwise)" gene 3705..4499 /gene="aph(3')-II" CDS 3705..4499 /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 4673..4803 /note="SV40 early polyA" promoter complement(4892..4921) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)" rep_origin complement(5245..5833) /note="pUC19 origin of replication () (RNAII -35 to RNA/DNA switch point) (old sequence errors)" gene complement(6004..6864) /gene="bla" CDS complement(6004..6864) /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 1679 a 1794 c 1835 g 1692 t ORIGIN 1 gacggatcgg gagatctaaa tgagtcttcg gacctcgcgg gggccgctta agcggtggtt 61 agggtttgtc tgacgcgggg ggagggggaa ggaacgaaac actctcattc ggaggcggct 121 cggggtttgg tcttggtggc cacgggcacg cagaagagcg ccgcgatcct cttaagcacc 181 cccccgccct ccgtggaggc gggggtttgg tcggcgggtg gtaactggcg ggccgctgac 241 tcgggcgggt cgcgcgcccc agagtgtgac cttttcggtc tgctcgcaga cccccgggcg 301 gcgccgccgc ggcggcgacg ggctcgctgg gtcctaggct ccatggggac cgtatacgtg 361 gacaggctct ggagcatccg cacgactgcg gtgatattac cggagacctt ctgcgggacg 421 agccgggtca cgcggctgac gcggagcgtc cgttgggcga caaacaccag gacggggcac 481 aggtacacta tcttgtcacc cggaggcgcg agggactgca ggagcttcag ggagtggcgc 541 agctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata 601 tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt gtcattggcg 661 aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc gcatattaag 721 gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa aagcttggta 781 ccgagctcgg atccgccacc atgaagacct taattcttgc cgttgcatta gtctactgcg 841 ccactgttca ttgccaggac tgtccttacg aacctgatcc accaaacaca gttccaactt 901 cctgtgaagc taaagaagga gaatgtattg atagcagctg tggcacctgc acgagagaca 961 tactatcaga tggactgtgt gaaaataaac caggaaaaac atgttgccga atgtgtcagt 1021 atgtaattga atgcagagta gaggccgcag gatggtttag aacattctat ggaaagagat 1081 tccagttcca ggaacctggt acatacgtgt tgggtcaagg aaccaagggc ggcgactgga 1141 aggtgtccat caccctggag aacctggatg gaaccaaggg ggctgtgctg accaagacaa 1201 gactggaagt ggctggagac atcattgaca tcgctcaagc tactgagaat cccatcactg 1261 taaacggtgg agctgaccct atcatcgcca acccgtacac catcggcgag gtcaccatcg 1321 ctgttgttga gatgccaggc ttcaacatca ccgtcattga gttcttcaaa ctgatcgtga 1381 tcgacatcct cggaggaaga tctgtaagaa tcgccccaga cacagcaaac aaaggaatga 1441 tctctggcct ctgtggagat cttaaaatga tggaagatac agacttcact tcagatccag 1501 aacaactcgc tattcagcct aagatcaacc aggagtttga cggttgtcca ctctatggaa 1561 atcctgatga cgttgcatac tgcaaaggtc ttctggagcc gtacaaggac agctgccgca 1621 accccatcaa cttctactac tacaccatct cctgcgcctt cgcccgctgt atgggtggag 1681 acgagcgagc ctcacacgtg ctgcttgact acagggagac gtgcgctgct cccgaaacta 1741 gaggaacctg cgttttgtct ggacatactt tctacgatac atttgacaaa gcaagatacc 1801 aattccaggg tccctgcaag gagattctta tggccgccga ctgtttctgg aacacttggg 1861 atgtgaaggt ttcacacagg aatgttgact cttacactga agtagagaaa gtacgaatca 1921 ggaaacaatc gactgtagta gaactcattg ttgatggaaa acagattctg gttggaggag 1981 aagccgtgtc cgtcccgtac agctctcaga acacttccat ctactggcaa gatggtgaca 2041 tactgactac agccatccta cctgaagctc tggtggtcaa gttcaacttc aagcaactgc 2101 tcgtcgtaca tattagagat ccattcgatg gtaagacttg cggtatttgc ggtaactaca 2161 accaggattt cagtgatgat tcttttgatg ctgaaggagc ctgtgatctg acccccaacc 2221 caccgggatg caccgaagaa cagaaacctg aagctgaacg actctgcaat agtctcttcg 2281 ccggtcaaag tgatcttgat cagaaatgta acgtgtgcca caagcctgac cgtgtcgaac 2341 gatgcatgta cgagtattgc ctgaggggac aacagggttt ctgtgaccac gcatgggagt 2401 tcaagaaaga atgctacata aagcatggag acaccctaga agtaccagat gaatgcaaat 2461 aggcggccgc accggtctcg agcatgcatc tagattcgag cagacatgat aagatacatt 2521 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt 2581 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac 2641 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag 2701 taaaacctct acaaatgtgg taaaatcgat aaactagaaa taattcttac tgtcatgcca 2761 agtaagatgc ttttctgtgc tgcaatagca ggcatgctgg ggatgcggtg ggctctatgg 2821 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg 2881 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 2941 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 3001 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc 3061 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga 3121 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 3181 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga 3241 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct 3301 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta 3361 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3421 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3481 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3541 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3601 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3661 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3721 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3781 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3841 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3901 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3961 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 4021 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 4081 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 4141 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 4201 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 4261 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 4321 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 4381 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4441 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4501 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 4561 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 4621 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 4681 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4741 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 4801 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 4861 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 4921 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 4981 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 5041 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 5101 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5161 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 5221 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 5281 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 5341 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 5401 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct 5461 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 5521 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 5581 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 5641 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat 5701 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 5761 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 5821 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 5881 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 5941 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 6001 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 6061 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 6121 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 6181 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 6241 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 6301 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 6361 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 6421 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 6481 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 6541 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 6601 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 6661 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 6721 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 6781 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 6841 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 6901 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 6961 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc //