Hello,
I think you are having an issue with the Anonymous Authentication Credentials on your website. Try this:
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Open IIS Manager
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Select your website
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Open the "Authentication" pane
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Select "Anonymous Authentication" and click the edit link in the Actions list.
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Change the "Anonymous user identity" to "Application pool identity"
Alternatively, you could grant permissions on your web directory to the user that is specified in the "Specific User" field. I find it easier to just use one identity for a website.
HTH,
Joe D.