LOCUS pNEBR-X1Hygro 6268 bp DNA circular 19-OCT-2007 DEFINITION Cloning vector pNEBR-X1Hygro, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pNEBR-X1Hygro ORGANISM Cloning vector pNEBR-X1Hygro other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6268) AUTHORS Maina,C.V. TITLE Direct Submission JOURNAL Submitted (19-OCT-2007) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6268 /organism="Cloning vector pNEBR-X1Hygro" /mol_type="other DNA" polyA_signal 4..855 /note="SV40 polyA" gene 995..1855 /gene="bla" CDS 995..1855 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" gene complement(2635..3672) /gene="hpt" CDS complement(2635..3672) /gene="hpt" /note="hygR, hph, aphIV (confers resistance to hygromycin)" /codon_start=1 /product="hygromycin B phosphotransferase" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS TRPDREMGEAN" rep_origin 4498..5086 /note="pUC19 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" terminator 5148..5606 /note="SV40 transcription terminator; prevents read-through transcription of the expression cassette from sources upstream of 5XRE" gene 5716..6240 /gene="lacZalpha" CDS 5716..6240 /gene="lacZalpha" /codon_start=1 /product="beta-galactosidase alpha fragment" /translation="MTMITPSYGRSTVLRAEYCPPSGVLSSERSTVLRAEFCPPSGDS RGYIMGAVSYYQSSWARAIEAGWRQASLQDIWIHEFLIKAGLERPLAIAARHASPDVQ ACVDPLVKAYSESYYGLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPS QQLRSLNGEWRFAW" promoter 5743..5835 /gene="lacZalpha" /note="5X GAL4 response element (5XRE) (approx. transcript start 3908 clockwise)" TATA_signal 5854..5860 /gene="lacZalpha" /note="minimal TATA box for 5XRE" misc_feature 5891..6049 /gene="lacZalpha" /note="multiple cloning site (ApaI-StuI)" promoter complement(6048..6065) /note="T7 promoter (transcript start 6048 counter-clockwise)" BASE COUNT 1595 a 1566 c 1535 g 1572 t ORIGIN 1 gttctagagg atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact 61 acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa 121 actactgatt ctaattgttt gtgtatttta gattccaacc tatggaactt atgaatggga 181 gcagtggtgg aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag 241 tgatgatgag gctactgctg actctcaaca ttctactctc aaaagaagag aaaggtagag 301 acccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt ttagtaatag 361 aactcttgct tgctttgcta tttacaacca caaaggaaaa agctgcactg ctatacaaga 421 aaattatgga aaaatattct gtaaccttta taagtaggca taacagttat aatcataaca 481 tactgttttt tcttactcca cacaggcata gagtgtctgc tattaataac tatgctcaaa 541 aattgtgtac ctttagcttt ttaatttgta aaggggttaa taaggaatat ttgatgtata 601 gtgccttgac tagagatcat aatcagccat accacatttg tagaggtttt acttgcttta 661 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt 721 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 781 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 841 tatcatgtct ggatcaacta cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc 901 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 961 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 1021 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 1081 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 1141 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttctccaa tgatgagcac 1201 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact 1261 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa 1321 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga 1381 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt 1441 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga 1501 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg 1561 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat 1621 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat 1681 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc 1741 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga 1801 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc 1861 agaccaagtt tactcatata tactttagat tgatttaccc cggttgataa tcagaaaagc 1921 cccaaaaaca ggaagattgt ataagcaaat attcatgttc tttcctgcgt tatcccctga 1981 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 2041 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct 2101 ccttacgcat ctgtgcggta tttcacaccg cactctctca tatgcgccag aaatccgcgc 2161 ggtggttttt gggggtcggg ggtgtttggc agccacagac gcccggtgtt cgtgtcgcgc 2221 cagtacatgc ggtccatgcc caggccatcc aaaaaccatg ggtctgtctg ctcagtccag 2281 tcgtggacct gaccccacgc aacgcccaaa ataataaccc ccacgaacca taaaccattc 2341 cccatggggg accccgtccc taacccacgg ggccagtggc tatggcaggg cctgccgccc 2401 cgacgttggc tgcgagccct gggccttcac ccgaacttgg ggggtggggt ggggaaaagg 2461 aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc 2521 cctgggaccg aaccccgcgt ttatgaacaa acgacccaac acccgtgcgt tttattctgt 2581 ctttttattg ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt 2641 agcctccccc atctcccgat ccggacgagt gctggggcgt cggtttccac tatcggcgag 2701 tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt gtgtacgccc 2761 gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc 2821 atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata 2881 cgcccggagc cgcggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg 2941 ctgctccata caagccaacc acggcctcca gaagaagatg ttggcgacct cgtattggga 3001 atccccgaac atcgcctcgc tccagtcaat gaccgctgtt atgcggccat tgtccgtcag 3061 gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc 3121 ccaaagcatc agctcatcga gagcctgcgc gacggacgca ctgacggtgt cgtccatcac 3181 agtttgccag tgatacacat ggggatcagc aatcgcgcat atgaaatcac gccatgtagt 3241 gtattgaccg attccttgcg gtccgaatgg gccgaacccg ctcgtctggc taagatcggc 3301 cgcagcgatc gcatccatgg cctccgcgac cggctgcaga acagcgggca gttcggtttc 3361 aggcaggtct tgcaacgtga caccctgtgc acggcgggag atgcaatagg tcaggctctc 3421 gctgaattcc ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg caaagtgccg 3481 ataaacataa cgatctttgt agaaaccatc ggcgcagcta tttacccgca ggacatatcc 3541 acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga gctgcatcag 3601 gtcggagacg ctgtcgaact tttcgatcag aaacttctcg acagacgtcg cggtgagttc 3661 aggctttttc atatcaagct gatctgcggc acgctgttga cgctgttaag cgggtcgctg 3721 cagggtcgct cggtgttcga ggccacacgc gtcaccttaa tatgcgaagt ggacctcgga 3781 ccgcgccgcc ccgactgcat ctgcgtgttc gaattcgcca atgacaagac gctgggcggg 3841 gtttgtgtca tcatagaact aaagacatgc aaatatattt cttcagggga caccgccagc 3901 aaacgcgagc aacgggccac ggggatgaag cagtaaaatt gtaaacgtta atattttgtt 3961 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg 4021 caaaatccct tataaatcaa aagaatagcc cgagataggg ttgagtgttg ttccagtttg 4081 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta 4141 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg 4201 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa 4261 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca 4321 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag 4381 ggcgcgtaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 4441 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 4501 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 4561 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 4621 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa 4681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 4741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 4801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 4861 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 4921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 4981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 5041 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 5101 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtatc agccatatca 5161 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac 5221 ataaaatgaa tgcaattgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa 5281 gcaatagcat cacaaatttc acaaataaag catttttcac tgcattctag ttgtggtttg 5341 tccaaactca tcaatgtatc ttatcatgtc tggatcagcc atatcacatt tgtagaggtt 5401 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 5461 attgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 5521 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 5581 aatgtatctt atcatgtctg ccatgtaatg tgagttagct cactcattag gcaccccagg 5641 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc 5701 acacaggaaa cagctatgac catgattacg ccaagctacg ggcggagtac tgtcctccga 5761 gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg 5821 gagttctgtc ctccgagcgg agactctaga gggtatataa tgggggccgt cagctactac 5881 cagagctcat gggcccgtgc aattgaagcc ggctggcgcc aagcttctct gcaggatatc 5941 tggatccacg aattcttaat taaggccggc ctcgagcggc cgctagcgat cgcggcgcgc 6001 cacgcgtctc cggatgtaca ggcatgcgtc gaccctctag tcaaggccta tagtgagtcg 6061 tattacggac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa 6121 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc 6181 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt cgcttggtaa 6241 taaagcccgc ttcggcgggc tttttttt //