LOCUS pCLuc-Basic2 6061 bp DNA circular 24-NOV-2009 DEFINITION Cloning vector pCLuc-Basic2, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pCLuc-Basic2 ORGANISM Cloning vector pCLuc-Basic2 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6061) AUTHORS Shea,C. and Tzertzinis,G. TITLE Direct Submission JOURNAL Submitted (24-NOV-2009) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6061 /organism="Cloning vector pCLuc-Basic2" /mol_type="other DNA" misc_feature 20..68 /note="multiple cloning site (EcoRI-BamHI)" misc_signal 69..78 /gene="CLuc" /note="Kozak consensus sequence" gene 75..1736 /gene="CLuc" CDS 75..1736 /gene="CLuc" /note="contains 30 codon substitutions for efficient mammalian translation" /codon_start=1 /product="Cypridina noctiluca luciferase" /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECID SSCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTY VLGQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADP IIANPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLC GDLKMMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPI NFYYYTISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQ FQGPCKEILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVG GEAVSVPYSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGIC GNYNQDFSDDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHK PDRVERCMYEYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK" sig_peptide 75..128 /gene="CLuc" /note="General Secretory Pathway signal peptide" polyA_signal 1745..1793 /gene="CLuc" /note="synthetic polyA" promoter 2379..2714 /gene="aph(3')-II" /note="SV40 promoter (clockwise)" gene 2766..3560 /gene="aph(3')-II" CDS 2766..3560 /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 3734..3855 /gene="aph(3')-II" /note="SV40 early polyA" rep_origin complement(4306..4894) /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5065..5925) /gene="bla" CDS complement(5065..5925) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1479 a 1547 c 1528 g 1507 t ORIGIN 1 gacggatcgg gagatcttgg aattctgcag atatcctcga gcccaagctt ggtaccgagc 61 tcggatccgc caccatgaag accttaattc ttgccgttgc attagtctac tgcgccactg 121 ttcattgcca ggactgtcct tacgaacctg atccaccaaa cacagttcca acttcctgtg 181 aagctaaaga aggagaatgt attgatagca gctgtggcac ctgcacgaga gacatactat 241 cagatggact gtgtgaaaat aaaccaggaa aaacatgttg ccgaatgtgt cagtatgtaa 301 ttgaatgcag agtagaggcc gcaggatggt ttagaacatt ctatggaaag agattccagt 361 tccaggaacc tggtacatac gtgttgggtc aaggaaccaa gggcggcgac tggaaggtgt 421 ccatcaccct ggagaacctg gatggaacca agggggctgt gctgaccaag acaagactgg 481 aagtggctgg agacatcatt gacatcgctc aagctactga gaatcccatc actgtaaacg 541 gtggagctga ccctatcatc gccaacccgt acaccatcgg cgaggtcacc atcgctgttg 601 ttgagatgcc aggcttcaac atcaccgtca ttgagttctt caaactgatc gtgatcgaca 661 tcctcggagg aagatctgta agaatcgccc cagacacagc aaacaaagga atgatctctg 721 gcctctgtgg agatcttaaa atgatggaag atacagactt cacttcagat ccagaacaac 781 tcgctattca gcctaagatc aaccaggagt ttgacggttg tccactctat ggaaatcctg 841 atgacgttgc atactgcaaa ggtcttctgg agccgtacaa ggacagctgc cgcaacccca 901 tcaacttcta ctactacacc atctcctgcg ccttcgcccg ctgtatgggt ggagacgagc 961 gagcctcaca cgtgctgctt gactacaggg agacgtgcgc tgctcccgaa actagaggaa 1021 cctgcgtttt gtctggacat actttctacg atacatttga caaagcaaga taccaattcc 1081 agggtccctg caaggagatt cttatggccg ccgactgttt ctggaacact tgggatgtga 1141 aggtttcaca caggaatgtt gactcttaca ctgaagtaga gaaagtacga atcaggaaac 1201 aatcgactgt agtagaactc attgttgatg gaaaacagat tctggttgga ggagaagccg 1261 tgtccgtccc gtacagctct cagaacactt ccatctactg gcaagatggt gacatactga 1321 ctacagccat cctacctgaa gctctggtgg tcaagttcaa cttcaagcaa ctgctcgtcg 1381 tacatattag agatccattc gatggtaaga cttgcggtat ttgcggtaac tacaaccagg 1441 atttcagtga tgattctttt gatgctgaag gagcctgtga tctgaccccc aacccaccgg 1501 gatgcaccga agaacagaaa cctgaagctg aacgactctg caatagtctc ttcgccggtc 1561 aaagtgatct tgatcagaaa tgtaacgtgt gccacaagcc tgaccgtgtc gaacgatgca 1621 tgtacgagta ttgcctgagg ggacaacagg gtttctgtga ccacgcatgg gagttcaaga 1681 aagaatgcta cataaagcat ggagacaccc tagaagtacc agatgaatgc aaataggcgg 1741 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgtctagaa 1801 ataattctta ctgtcatgcc aagtaagatg cttttctgtg ctgcaatagc aggcatgctg 1861 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt 1921 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 1981 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 2041 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc 2101 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 2161 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 2221 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 2281 atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa 2341 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 2401 ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 2461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 2521 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 2581 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg 2641 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 2701 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt 2761 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 2821 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 2881 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 2941 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 3001 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 3061 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 3121 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 3181 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 3241 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc 3301 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 3361 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 3421 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 3481 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 3541 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc 3601 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 3661 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 3721 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 3781 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 3841 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc 3901 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 3961 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 4021 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 4081 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 4141 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 4201 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 4261 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 4321 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 4381 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 4441 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 4501 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 4561 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 4621 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 4681 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 4741 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 4801 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 4861 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 4921 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 4981 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5041 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5101 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5161 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 5221 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 5281 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 5341 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 5401 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 5461 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 5521 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 5581 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 5641 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 5701 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 5761 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 5821 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 5881 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 5941 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6001 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6061 c //