LOCUS pCLuc-SP6 6902 bp DNA circular 16-DEC-2016 DEFINITION Cloning vector pCLuc-SP6, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pCLuc-SP6 ORGANISM Cloning vector pCLuc-SP6 Unclassified. REFERENCE 1 (bases 1 to 6902) AUTHORS Schwarz,D. and Schildkraut,E. TITLE Direct Submission JOURNAL Submitted (16-DEC-2016) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6902 /organism="Cloning vector pCLuc-SP6" /mol_type="other DNA" regulatory 209..863 /regulatory_class="promoter" /note="CMV promoter (clockwise)" regulatory 863..879 /regulatory_class="promoter" /note="SP6 promoter" misc_RNA 911..920 /note="Kozak consensus sequence for CLuc" gene 917..2578 /gene="CLuc" CDS 917..2578 /gene="CLuc" /codon_start=1 /product="Cypridina noctiluca luciferase" /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECID SSCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTY VLGQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADP IIANPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLC GDLKMMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPI NFYYYTISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQ FQGPCKEILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVG GEAVSVPYSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGIC GNYNQDFSDDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHK PDRVERCMYEYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK" misc_feature 2636..2641 /note="XbaI restriction site; plasmid supplied linearized, cleaved between 2636-2637" misc_feature 2637 /note="5' end of linearized plasmid" regulatory 3221..3556 /regulatory_class="promoter" /note="SV40 promoter (clockwise)" gene 3607..4401 /gene="aph(3')-II" CDS 3607..4401 /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" regulatory complement(4794..4824) /regulatory_class="promoter" /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)" rep_origin complement(5147..5735) /note="pUC19 origin of replication () (RNAII -35 to RNA/DNA switch point) (old sequence errors)" gene complement(5906..6766) /gene="bla" CDS complement(5906..6766) /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 1689 a 1751 c 1729 g 1733 t ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa atatttaggt gacactatag aagcccaagc ttggtaccga 901 gctcggatcc gccaccatga agaccttaat tcttgccgtt gcattagtct actgcgccac 961 tgttcattgc caggactgtc cttacgaacc tgatccacca aacacagttc caacttcctg 1021 tgaagctaaa gaaggagaat gtattgatag cagctgtggc acctgcacga gagacatact 1081 atcagatgga ctgtgtgaaa ataaaccagg aaaaacatgt tgccgaatgt gtcagtatgt 1141 aattgaatgc agagtagagg ccgcaggatg gtttagaaca ttctatggaa agagattcca 1201 gttccaggaa cctggtacat acgtgttggg tcaaggaacc aagggcggcg actggaaggt 1261 gtccatcacc ctggagaacc tggatggaac caagggggct gtgctgacca agacaagact 1321 ggaagtggct ggagacatca ttgacatcgc tcaagctact gagaatccca tcactgtaaa 1381 cggtggagct gaccctatca tcgccaaccc gtacaccatc ggcgaggtca ccatcgctgt 1441 tgttgagatg ccaggcttca acatcaccgt cattgagttc ttcaaactga tcgtgatcga 1501 catcctcgga ggaagatctg taagaatcgc cccagacaca gcaaacaaag gaatgatctc 1561 tggcctctgt ggagatctta aaatgatgga agatacagac ttcacttcag atccagaaca 1621 actcgctatt cagcctaaga tcaaccagga gtttgacggt tgtccactct atggaaatcc 1681 tgatgacgtt gcatactgca aaggtcttct ggagccgtac aaggacagct gccgcaaccc 1741 catcaacttc tactactaca ccatctcctg cgccttcgcc cgctgtatgg gtggagacga 1801 gcgagcctca cacgtgctgc ttgactacag ggagacgtgc gctgctcccg aaactagagg 1861 aacctgcgtt ttgtctggac atactttcta cgatacattt gacaaagcaa gataccaatt 1921 ccagggtccc tgcaaggaga ttcttatggc cgccgactgt ttctggaaca cttgggatgt 1981 gaaggtttca cacaggaatg ttgactctta cactgaagta gagaaagtac gaatcaggaa 2041 acaatcgact gtagtagaac tcattgttga tggaaaacag attctggttg gaggagaagc 2101 cgtgtccgtc ccgtacagct ctcagaacac ttccatctac tggcaagatg gtgacatact 2161 gactacagcc atcctacctg aagctctggt ggtcaagttc aacttcaagc aactgctcgt 2221 cgtacatatt agagatccat tcgatggtaa gacttgcggt atttgcggta actacaacca 2281 ggatttcagt gatgattctt ttgatgctga aggagcctgt gatctgaccc ccaacccacc 2341 gggatgcacc gaagaacaga aacctgaagc tgaacgactc tgcaatagtc tcttcgccgg 2401 tcaaagtgat cttgatcaga aatgtaacgt gtgccacaag cctgaccgtg tcgaacgatg 2461 catgtacgag tattgcctga ggggacaaca gggtttctgt gaccacgcat gggagttcaa 2521 gaaagaatgc tacataaagc atggagacac cctagaagta ccagatgaat gcaaataggc 2581 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgtctag 2641 aaataattct tactgtcatg ccaagtaaga tgcttttctg tgctgcaata gcaggcatgc 2701 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg 2761 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag 2821 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt 2881 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt 2941 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg 3001 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 3061 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 3121 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 3181 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 3241 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 3301 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3361 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 3421 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct 3481 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 3541 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg 3601 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg 3661 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg 3721 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 3781 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 3841 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 3901 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 3961 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 4021 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 4081 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg 4141 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 4201 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat 4261 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 4321 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 4381 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc 4441 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg 4501 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt 4561 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 4621 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 4681 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat 4741 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 4801 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 4861 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 4921 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 4981 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 5041 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 5101 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 5161 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 5221 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 5281 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 5341 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 5401 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 5461 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 5521 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 5581 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 5641 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 5701 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 5761 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 5821 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 5881 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 5941 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 6001 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 6061 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 6121 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 6181 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 6241 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 6301 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 6361 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 6421 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 6481 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 6541 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 6601 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 6661 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 6721 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 6781 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 6841 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 6901 tc //