Product Data Sheet

Map of the plasmid:

Recovery

1. Obtain an LB agar plate with the appropriate antibiotic.

2. Using a sterile pipette tip, touch the bacteria growing within the punctured area of the stab culture. (A sterilized wire loop or sterile toothpick can be used in place of a sterile pipette tip.)

3. Run this tip lightly over a section of the plate, as shown in the figure, to create streak #1.

4. Using another sterile pipette tip, pass through streak #1 and spread the bacteria over a second section of the plate, to create streak #2.

5. Using a third sterile pipette tip, pass through streak #2 and spread the bacteria over the last section of the plate, to create streak #3.

6. Grow overnight in a 37 ^o C incubator (unless a different growth temperature is indicated on the plasmid datasheet).

7. In the morning, single colonies should be visible. If the bacterial growth is too dense, re-streak onto a new agar plate to obtain single colonies.

载体序列：

LOCUS       pAN7-1                  6756 bp    DNA     circular   SYN 14-NOV-2006

DEFINITION  Expression vector (PAN7-1) DNA, 6756bp.

ACCESSION   Z32698

VERSION     Z32698.1  GI:475166

KEYWORDS    hygromycin b phosphotransferase.

SOURCE      synthetic construct

  ORGANISM  synthetic construct

            other sequences; artificial sequences.

COMMENT     This file is created by Vector NTI

            http://www.biofeng.com/

COMMENT     VNTAUTHORNAME|biofeng.com|

REFERENCE   1

  AUTHORS   Mullaney,E.J., Hamer,J.E., Roberti,K.A., Yelton,M.M. and

            Timberlake,W.E.

  TITLE     Primary structure of the trpC gene from Aspergillus nidulans

  JOURNAL   Mol. Gen. Genet. 199 (1), 37-45 (1985)

   PUBMED   3158796

  REMARK    sites .

FEATURES             Location/Qualifiers

     source          1..6756

                     /organism="synthetic construct"

                     /mol_type="other DNA"

                     /db_xref="taxon:32630"

     misc_feature    1..6756

                     /product="fungal transformation vector pAN7-1"

                     /standard_name="HPH"

                     /experiment="experimental evidence, no additional details

                     recorded"

                     /citation=[2]

                     /phenotype="gene product confers resistance to hygromycin

                     B"

     promoter        1..2129

                     /experiment="experimental evidence, no additional details

                     recorded"

                     /citation=[2]

                     /citation=[3]

                     /function="promoter region of the A.nidulans gpdA gene"

     gene            2130

                     /gene="gpdA"

     misc_feature    2130

                     /gene="gpdA"

                     /experiment="experimental evidence, no additional details

                     recorded"

                     /citation=[3]

                     /function="major transcription start site"

     intron          2178..2293

                     /experiment="experimental evidence, no additional details

                     recorded"

                     /citation=[3]

                     /number=1

     gene            2302..3321

                     /gene="HPH"

     CDS             2302..3321

                     /gene="HPH"

                     /experiment="experimental evidence, no additional details

                     recorded"

                     /label="gene-product confers resistance to hygromycin B"

                     /citation=[2]

                     /citation=[3]

                     /codon_start=1

                     /transl_table=11

                     /product="hygromycin B phosphotransferase"

                     /protein_id="CAA83647.1"

                     /db_xref="GI:475167"

                     /translation="MPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY

                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP

                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW

                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEA

                     MFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSL

                     VDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTR

                     PRAKE"

     terminator      3339..4108

                     /standard_name="terminator region from A.nidulans trpC

                     gene (containing CDS)"

                     /experiment="experimental evidence, no additional details

                     recorded"

                     /label="this region contains the 3end of the trpC

                     transcript including 50 codons. two major transcription

                     ends have been identified"

                     /citation=[1]

                     /citation=[2]

     polyA_site      3590..3600

     misc_difference 3600..3610

                     /label="introduction of one base at this position generates

                     an BLGI site with was shown to be present by restriction

                     digestion"

                     /citation=[1]

                     /replace="gccttncaggc"

     misc_difference 3809..3817

                     /label="introduction of one base at this position generates

                     an ALWNI site which was shown to be present by restriction

                     digestion"

                     /citation=[1]

                     /replace="cagcncctg"

     polyA_site      3863..3873

     misc_difference 3897..3907

                     /label="introduction (actual sequence could be deletion as

                     well) of a base at this position results in the

                     elimination of an XMNI site which was shown to be absent

                     by restriction digestion"

                     /citation=[1]

                     /replace="gaaatcanttc"

     variation       3934..3941

                     /label="A NCOI site present at this position was removed by

                     digestion, treatment with klenow polymerase and

                     religation"

                     /citation=[2]

                     /replace="catgcatg"

     misc_feature    4105..6756

                     /product="PUC18"

                     /label="vector sequences derived from PUC18 from SALI TO

                     ECORI"

                     /citation=[2]

ORIGIN      

        1 gaattccctt gtatctctac acacaggctc aaatcaataa gaagaacggt tcgtcttttt

       61 cgtttatatc ttgcatcgtc cc