LOCUS Exported 5802 bp ds-DNA circular SYN 03-8-2013 DEFINITION Control vector with a strong constitutive CMV promoter for expressing secreted Gaussia luciferase in mammalian cells. ACCESSION . VERSION . KEYWORDS pCMV-GLuc 2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5802) AUTHORS New England Biolabs TITLE Direct Submission FEATURES Location/Qualifiers source 1..5802 /organism="synthetic DNA construct" /lab_host="Mammalian Cells" /mol_type="other DNA" enhancer 235..614 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" promoter 863..881 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" misc_signal 914..923 /note="Kozak sequence" CDS 920..1477 /codon_start=1 /product="secreted Gaussia luciferase" /note="hGLuc" /note="human codon-optimized" /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ RCATFASKIQGQVDKIKGAGGD" polyA_signal 1486..1534 /note="synthetic polyadenylation signal" rep_origin 1668..2096 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2110..2440 /note="SV40 promoter" /note="SV40 enhancer and early promoter" rep_origin 2291..2426 /note="SV40 ori" /note="SV40 origin of replication" CDS 2507..3301 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /note="NeoR/KanR" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 3475..3596 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" rep_origin complement(4047..4635) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4806..5666) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5667..5771) /gene="bla" /note="AmpR promoter" 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 atgggtggac 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 attaatacga ctcactatag ggagacccaa gcttggtacc 901 gagctcggat ccagccacca tgggagtcaa agttctgttt gccctgatct gcatcgctgt 961 ggccgaggcc aagcccaccg agaacaacga agacttcaac atcgtggccg tggccagcaa 1021 cttcgcgacc acggatctcg atgctgaccg cgggaagttg cccggcaaga agctgccgct 1081 ggaggtgctc aaagagatgg aagccaatgc ccggaaagct ggctgcacca ggggctgtct 1141 gatctgcctg tcccacatca agtgcacgcc caagatgaag aagttcatcc caggacgctg 1201 ccacacctac gaaggcgaca aagagtccgc acagggcggc ataggcgagg cgatcgtcga 1261 cattcctgag attcctgggt tcaaggactt ggagcccatg gagcagttca tcgcacaggt 1321 cgatctgtgt gtggactgca caactggctg cctcaaaggg cttgccaacg tgcagtgttc 1381 tgacctgctc aagaagtggc tgccgcaacg ctgtgcgacc tttgccagca agatccaggg 1441 ccaggtggac aagatcaagg gggccggtgg tgactaagcg gccgcaataa aatatcttta 1501 ttttcattac atctgtgtgt tggttttttg tgtgtctaga aataattctt actgtcatgc 1561 caagtaagat gcttttctgt gctgcaatag caggcatgct ggggatgcgg tgggctctat 1621 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag 1681 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 1741 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 1801 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca 1861 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata 1921 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 1981 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg 2041 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt 2101 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag 2161 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 2221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 2281 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 2341 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa 2401 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat 2461 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 2521 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 2581 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 2641 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc 2701 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 2761 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 2821 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 2881 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 2941 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 3001 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt 3061 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 3121 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 3181 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 3241 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 3301 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 3361 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 3421 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg 3481 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 3541 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 3601 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct 3661 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 3721 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 3781 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 3841 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 3901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 3961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 4021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 4081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 4141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 4201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat 4261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 4321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 4381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 4441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 4501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 4561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 4621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 4681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 4741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 4801 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 4861 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 4921 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 4981 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 5041 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 5101 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 5161 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 5221 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 5281 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 5341 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 5401 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 5461 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 5521 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 5581 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 5641 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 5701 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 5761 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //