LOCUS PCRII 3932 bp DNA circular SYN 17-MAY-1996 DEFINITION pCRII. ACCESSION PCRII0 KEYWORDS . SOURCE synthetic construct. ORGANISM synthetic construct Unclassified. REFERENCE 1 (bases 1 to 3932) AUTHORS Fristensky,B. TITLE untitled JOURNAL Unpublished Plant Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada COMMENT PCR products generated by Taq polymerase typically contain a single 5' protruding A. To clone these products, pCRII is cut at the EcoRI site and an EcoRI adaptor is added to each end to give the following structure: 5' GAATTCGGCTT AATTC3' 3' CTTAA TTCGGCTTAAG5' The 5' overhanging A will base pair with the one of the T's, and presumably E. coli will repair the site. FEATURES Location/Qualifiers source 1..3932 /organism="synthetic construct" /plasmid="" rep_origin 572..986 /standard_name="F1" rep_origin 3182..3765 /standard_name="ColE1" promoter 239..255 /label=Sp6_promoter misc_feature 269..381 /label=MCS promoter 388..407 /label=T7_promoter misc_feature 1..571 /gene="lacZ" /note="beta-galactosidase~" misc_feature 987..2114 /gene="nptII" /note="Description: kanamycin resistance" misc_feature 2133..2992 /gene="AmpR" /note="Description: ampicillin resistance" BASE COUNT 915 a 1011 c 1065 g 941 t ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat 241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca 301 ctagtaacgg ccgccagtgt gctggaattc tgcagatatc catcacactg gcggccgctc 361 gagcatgcat ctagagggcc caattcgccc tatagtgagt cgtattacaa ttcactggcc 421 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca 481 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc 541 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg 601 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 661 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 721 aatcgggggc tccctttagg gttccgattt agagctttac ggcacctcga ccgcaaaaaa 781 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 841 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 901 aaccctatcg cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 961 ttaaaaaatg agctgattta acaaattcag ggcgcaaggg ctgctaaagg aaccggaaca 1021 cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta 1081 tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat 1141 ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg 1201 cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa 1261 ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatgagga tcgtttcgca 1321 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 1381 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 1441 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 1501 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 1561 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 1621 atctcctgtc atctcgcctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 1681 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 1741 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 1801 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 1861 gcgaggatct cgtcgtgatc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 1921 gccgcttttc tggattcaac gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 1981 tagcgttgga tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 2041 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 2101 acgagttctt ctgaattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct 2161 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa 2221 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa 2281 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt 2341 taaagttctg ctatgtgata cactattatc ccgtattgac gccgggcaag agcaactcgg 2401 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca 2461 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa 2521 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt 2581 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc 2641 cataccaaac gacgagagtg acaccacgat gcctgtagca atgccaacaa cgttgcgcaa 2701 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actgaatgga 2761 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc 2821 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga 2881 tggtaagcgc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga 2941 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga 3001 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat 3061 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt 3121 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 3181 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 3241 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 3301 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 3361 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 3421 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 3481 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 3541 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 3601 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 3661 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 3721 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 3781 cctgggcttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 3841 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga 3901 gcgcagcgag tcagtgagcg aggaagcgga ag //